Busquets, Nuria; Xavier, F; Martín-Folgar, Raquel; Lorenzo, Gema; Galindo-Cardiel, Iván; del Val, Bernat Pérez; Rivas, Raquel; Iglesias, Javier; Rodríguez, Fernando; Solanes, David; Domingo, Mariano; Brun, Alejandro
The increasing interest in Rift Valley fever virus (RVFV) and its potential impact on naive animal populations deserve revisiting experimental reproduction of RVFV infection, particularly in those animal breeds for which no data about their susceptibility to RVFV infection have ever been recorded. In this study we show the susceptibility of 9-10 weeks old European sheep (Ripollesa breed) to RVFV infection, showing a mild, subacute form of disease. Four different viral isolates efficiently replicated in vivo after subcutaneous experimental inoculation, and consistent viral loads in blood and virus shedding (variable in length depending on the RVFV isolate used) were detected, showing horizontal transmission to a noninfected, sentinel lamb. RVFV infection caused transient pyrexia in adult lambs and no other clinical symptoms were observed, with the exception of corneal opacity ("blue eye") found in 3 out of 16 subcutaneously inoculated sheep. In conclusion, adult sheep from this European breed are readily infected with RVFV without apparent clinical manifestations.
López-Olvera, Jorge Ramón; Falconi, Caterina; Férnandez-Pacheco, Paloma; Fernández-Pinero, Jovita; Sánchez, Miguel Angel; Palma, Agustín; Herruzo, Irene; Vicente, Joaquín; Jiménez-Clavero, Miguel Angel; Arias, Marisa; Sánchez-Vizcaíno, José Manuel; Gortázar, Christian
Bluetongue (BT) is a climate change-related emerging infectious disease in Europe. Outbreaks of serotypes 1, 2, 4, 6, 8, 9, 11, and 16 are challenging Central and Western Europe since 1998. Measures to control or eradicate bluetongue virus (BTV) from Europe have been implemented, including movement restrictions and vaccination of domestic BTV-susceptible ruminants. However, these measures are difficult to apply in wild free-ranging hosts of the virus, like red deer (Cervus elaphus), which could play a role in the still unclear epidemiology of BT in Europe. We show for the first time that BTV RNA can be detected in European red deer blood for long periods, comparable to those of domestic ruminants, after experimental infection with BTV-1 and BTV-8. BTV RNA was detected in experimentally infected red deer blood up to the end of the study (98-112 dpi). BTV-specific antibodies were found in serum both by enzyme-linked immunosorbent assay (ELISA) and virus neutralization (VNT) from 8 to 12 dpi to the end of the study, peaking at 17-28 dpi. Our results indicate that red deer can be infected with BTV and maintain BTV RNA for long periods, remaining essentially asymptomatic. Thus, unvaccinated red deer populations have the potential to be a BT reservoir in Europe, and could threaten the success of the European BTV control strategy. Therefore, wild and farmed red deer should be taken into account for BTV surveillance, and movement restrictions and vaccination schemes applied to domestic animals should be adapted to include farmed or translocated red deer.
Cliquet, Florence; Picard-Meyer, Evelyne; Barrat, Jacques; Brookes, Sharon M; Healy, Derek M; Wasniewski, Marine; Litaize, Estelle; Biarnais, Mélanie; Johnson, Linda; Fooks, Anthony R
Background Since 1954, there have been in excess of 800 cases of rabies as a result of European Bat Lyssaviruses types 1 and 2 (EBLV-1, EBLV-2) infection, mainly in Serotine and Myotis bats respectively. These viruses have rarely been reported to infect humans and terrestrial mammals, as the only exceptions are sheep in Denmark, a stone marten in Germany and a cat in France. The purpose of this study was to investigate the susceptibility of foxes to EBLVs using silver foxes (Vulpes vulpes) as a model. Results Our experimental studies have shown that the susceptibility of foxes to EBLVs is low by the intramuscular (IM) route, however, animals were sensitive to intracranial (IC) inoculation. Mortality was 100% for both EBLV-1 (~4.5 logs) and EBLV-2 (~3.0 logs) delivered by the IC route. Virus dissemination and inflammatory infiltrate in the brain were demonstrated but virus specific neutralising antibody (VNA) was limited (log(ED50) = 0.24–2.23 and 0.95–2.39 respectively for specific EBLV-1 and EBLV-2). Foxes were also susceptible, at a low level, to peripheral (IM) infection (~3.0 logs) with EBLV-1 but not EBLV-2. Three out of 21 (14.3%) foxes developed clinical signs between 14 and 24 days post-EBLV-1 infection. None of the animals given EBLV-2 developed clinical disease. Conclusion These data suggest that the chance of a EBLV spill-over from bat to fox is low, but with a greater probability for EBLV-1 than for EBLV-2 and that foxes seem to be able to clear the virus before it reaches the brain and cause a lethal infection. PMID:19454020
Millán, J; Casais, R; Colomar, V; Bach, E; Prieto, J M; Velarde, R
Scabies was recently reported for the first time in the European wild rabbit, Oryctolagus cuniculus (Lagomorpha: Leporidae). We experimentally exposed 10 seronegative wild-caught rabbits to skin from a mangy wild rabbit. Serological, physiological, parasitological and histopathological changes were recorded. Three rabbits developed antibodies at 2-5 weeks post-infection (w.p.i.), two of which then developed lesions at 7 w.p.i. One of these had a small area of alopecia on the hind limb that healed naturally within 1 week; the other developed more extensive lesions restricted to the hind limbs (as typically observed in wild rabbits) that lasted until the rabbit died (12.5 w.p.i.). The third rabbit died of trauma 5 w.p.i. before developing any lesions. Antibodies in the healed rabbit disappeared from serum at 8 w.p.i., whereas antibody levels in the sick rabbit increased until its death. Disseminated intravascular coagulation and hepatic necrosis, probably arising from a concomitant infection with rabbit haemorrhagic disease virus, were the likely final cause of death in this rabbit. The mangy rabbit that served as a donor died of a multifocal fibrinosuppurative pneumonia that may have been secondary to the skin bacterial pyoderma.
Drolet, Barbara S; Reister, Lindsey M; Rigg, Tara D; Nol, Pauline; Podell, Brendan K; Mecham, James O; VerCauteren, Kurt C; van Rijn, Piet A; Wilson, William C; Bowen, Richard A
Bluetongue (BT) is an insect-transmitted, economically important disease of domestic and wild ruminants. Although only five of the 26 reported bluetongue virus (BTV) serotypes are considered endemic to the USA, 10 exotic serotypes have been isolated primarily in the southeastern region of the country since 1999. For an exotic BTV serotype to become endemic there must be susceptible animal species and competent vectors. In the USA, sheep and white-tailed deer (WTD) are the primary sentinel livestock and wildlife species, respectively. In 2006, BTV-8 was introduced into Northern Europe and subsequently overwintered, causing unprecedented livestock disease and mortality during the 2006-2007 vector seasons. To assess the risk of the European strain of BTV-8 to North American WTD, and understand the role they could play after a similar introduction, eight bluetongue-seronegative WTD were inoculated with BTV-8. Body temperatures and clinical signs were recorded daily. Blood samples were analyzed for BTV RNA with quantitative real time reverse transcriptase polymerase chain reaction (qRT-PCR), serum analyzed for BTV antibodies by cELISA, and tissues taken for histopathology and qRT-PCR. All eight deer became infected and developed moderate to severe clinical disease from days 8 to 15. Peak viremia was from day 7 to 10 with detectable titers through the end of the study (28 days) in most deer. Serum antibody was detected by day 6, peaked by day 10 and continued through day 28. We conclude that North American WTD are highly susceptible to BTV-8 and would act as clinical disease sentinels and amplifying hosts during an outbreak.
Vendramin, N; Toffan, A; Mancin, M; Cappellozza, E; Panzarin, V; Bovo, G; Cattoli, G; Capua, I; Terregino, C
Viral encephalopathy and retinopathy (VER), otherwise known as viral nervous necrosis (VNN), is a severe pathological condition caused by RNA viruses belonging to the Nodaviridae family, genus Betanodavirus. The disease, described in more than 50 fish species worldwide, is considered as the most serious viral threat affecting marine farmed species in the Mediterranean region, thus representing one of the bottlenecks for further development of the aquaculture industry. To date, four different genotypes have been identified, namely red-spotted grouper nervous necrosis virus (RGNNV), striped jack nervous necrosis virus (SJNNV), tiger puffer nervous necrosis virus and barfin flounder nervous necrosis virus, with the RGNNV genotype appearing as the most widespread in the Mediterranean region, although SJNNV-type strains and reassortant viruses have also been reported. The existence of these genetically different strains could be the reason for the differences in mortality observed in the field. However, very little experimental data are available on the pathogenicity of these viruses in farmed fish. Therefore, in this study, the pathogenicity of 10 isolates has been assessed with an in vivo trial. The investigation was conducted using the European sea bass, the first target fish species for the disease in the Mediterranean basin. Naive fish were challenged by immersion and clinical signs and mortality were recorded for 68 days; furthermore, samples collected at selected time points were analysed to evaluate the development of the infection. Finally, survivors were weighed to estimate the growth reduction. The statistically supported results obtained in this study demonstrated different pathogenicity patterns, underlined the potential risk represented by different strains in the transmission of the infection to highly susceptible species and highlighted the indirect damage caused by a clinical outbreak of VER/VNN.
Corner, L A L; Costello, E; Lesellier, S; O'Meara, D; Gormley, E
The aim of the study was to describe, over a period of 24 weeks, the pathological and bacteriological changes in badgers experimentally infected with Mycobacterium bovis. The badgers were infected by endobronchial instillation of 2.5 x10(4) colony forming units (cfu) M. bovis. After infection, the badgers were examined at 3 weekly intervals when blood and tracheal aspirates were collected. At 6, 12, 18 and 24 weeks post-infection (pi) three animals were euthanized and a detailed pathological and bacteriological examination was performed to assess the nature of the experimental disease. During the course of the study only one badger developed clinical signs of disease: a subcutaneous swelling on its head, first observed at 18 weeks pi. At post-mortem examination gross and histological lesions of tuberculosis were observed and M. bovis was recovered from all, except one badger. In the majority of badgers the endobronchial route of inoculation resulted in the establishment of infection that over 24 weeks was non-progressive with limited dissemination of infection from the thoracic cavity, mainly to the hepatic and mesenteric lymph nodes. However, in one of the badgers examined at 18 weeks pi and one at 24 weeks pi, infection was widely disseminated. The disease induced by the endobronchial inoculation displayed the characteristics of disease observed in naturally infected badgers.
Lim, Stephanie M.; Brault, Aaron C.; van Amerongen, Geert; Sewbalaksing, Varsha D.; Osterhaus, Albert D. M. E.; Martina, Byron E. E.; Koraka, Penelope
Mass bird mortality has been observed in North America after the introduction of West Nile virus (WNV), most notably massive die-offs of American crows (Corvus brachyrhynchos). In contrast, WNV epidemic activity in Europe has been characterized by very low incidences of bird mortality. As the general susceptibility of European corvids to strains of WNV remains in question, European jackdaws (Corvus monedula) were inoculated with WNV strains circulating currently in Greece (Greece-10), Italy (FIN and Ita09) and Hungary (578/10), as well as a North American (NY99) genotype with a demonstrated corvid virulence phenotype. Infection with all strains except WNV-FIN resulted in mortality. Viraemia was observed for birds inoculated with all strains and virus was detected in a series of organs upon necropsy. These results suggested that jackdaws could potentially function as a sentinel for following WNV transmission in Europe; however, elicited viraemia levels might be too low to allow for efficient transmission of virus to mosquitoes. PMID:24671752
Busquets, Núria; Segalés, Joaquim; Córdoba, Lorena; Mussá, Tufaria; Crisci, Elisa; Martín-Valls, Gerard E; Simon-Grifé, Meritxell; Pérez-Simó, Marta; Pérez-Maíllo, Monica; Núñez, Jose I; Abad, Francesc X; Fraile, Lorenzo; Pina, Sonia; Majó, Natalia; Bensaid, Albert; Domingo, Mariano; Montoya, María
The recent pandemic caused by human influenza virus A(H1N1) 2009 contains ancestral gene segments from North American and Eurasian swine lineages as well as from avian and human influenza lineages. The emergence of this A(H1N1) 2009 poses a potential global threat for human health and the fact that it can infect other species, like pigs, favours a possible encounter with other influenza viruses circulating in swine herds. In Europe, H1N1, H1N2 and H3N2 subtypes of swine influenza virus currently have a high prevalence in commercial farms. To better assess the risk posed by the A(H1N1) 2009 in the actual situation of swine farms, we sought to analyze whether a previous infection with a circulating European avian-like swine A/Swine/Spain/53207/2004 (H1N1) influenza virus (hereafter referred to as SwH1N1) generated or not cross-protective immunity against a subsequent infection with the new human pandemic A/Catalonia/63/2009 (H1N1) influenza virus (hereafter referred to as pH1N1) 21 days apart. Pigs infected only with pH1N1 had mild to moderate pathological findings, consisting on broncho-interstitial pneumonia. However, pigs inoculated with SwH1N1 virus and subsequently infected with pH1N1 had very mild lung lesions, apparently attributed to the remaining lesions caused by SwH1N1 infection. These later pigs also exhibited boosted levels of specific antibodies. Finally, animals firstly infected with SwH1N1 virus and latter infected with pH1N1 exhibited undetectable viral RNA load in nasal swabs and lungs after challenge with pH1N1, indicating a cross-protective effect between both strains.
Tornos, Pilar; Gonzalez-Alujas, Teresa; Thuny, Frank; Habib, Gilbert
Infective endocarditis (IE) is a difficult and complex disease. In recent years epidemiology and microbiology have changed. In developed countries IE is now affecting older patients and patients with no previously known valve disease. Prosthetic IE (prosthetic valve endocarditis [PVE]) and endocarditis in patients with pacemakers and other devices (cardiac device related infective endocarditis [CDRIE]) are becoming more frequent. The number of Staphylococcus aureus IE is increasing related to the number of endocarditis that occurs because of health care associated procedures, especially in diabetics or patients on chronic hemodialysis. The change in the underlying population and the increase in the number of cases caused by very virulent organism explain why the disease still carries a poor prognosis and a high mortality. The variety of clinical manifestations and complications, as well as the serious prognosis, makes it mandatory that IE patients need to be treated in experienced hospitals with a collaborative approach between different specialists, involving cardiologists, infectious disease specialists, microbiologists, surgeons, and frequently others, including neurologists and radiologists. Only an early diagnosis followed by risk stratification and a prompt institution of the correct antibiotic treatment as well as an appropriate and timed surgical indication may improve mortality figures. The recent European Guidelines try to provide clear and simple recommendations, obtained by expert consensus after thorough review of the available literature to all specialists involved in clinical decision-making of this difficult and changing disease.
Kapel, C M O; Measures, L; Møller, L N; Forbes, L; Gajadhar, A
The susceptibility of seals to infection with Trichinella nativa and the cold tolerant characteristics of muscle larvae in seal meat were evaluated. Two grey seals, Halichoerus grypus, were inoculated with 5000 (100 larvae/kg) T. nativa larvae and two grey seals with 50000 (1000 larvae/kg). One seal from each dose group and two control seals were killed at 5 and 10 weeks post-inoculation (p.i.). At 5 weeks p.i., infection was established in both low and high dose seals with mean larval densities of 68 and 472 larvae per gram (lpg), respectively, using eight different muscles for analyses. At 10 weeks p.i., mean larval densities were 531 and 2649 lpg, respectively, suggesting an extended persistence of intestinal worms. In seals with high larval density infections, the distribution of larvae in various muscles was uniform, but in one seal with a low larval density infection, predilection sites of larvae included muscle groups with a relative high blood flow, i.e. diaphragm, intercostal and rear flipper muscles. Trichinella-specific antibody levels, as measured by ELISA, increased during the 10 week experimental period. Infected seal muscle was stored at 5, -5 and -18 degrees C for 1, 4 and 8 weeks. Muscle larvae released from stored seal muscle by artificial digestion were inoculated into mice to assess viability and infectivity. Larvae from seal muscle 10 weeks p.i. tolerated -18 degrees C for 8 weeks but larvae from seal muscle 5 weeks p.i. tolerated only 1 week at -18 degrees C, supporting the hypothesis that freeze tolerance increases with the age of the host-parasite tissue complex. The expressed susceptibility to infection, extended production of larvae, antibody response and freeze tolerance of T. nativa in seals are new findings from the first experimental Trichinella infection in any marine mammal and suggest that pinnipeds (phocids, otariiids or walrus) may acquire Trichinella infection by scavenging even small amounts of infected tissue left by hunters or
Potgieter, L N; Jones, J B; Patton, C S; Webb-Martin, T A
Five eight week old dogs were inoculated orally and intranasally with cell culture origin canine parvovirus. Three dogs became depressed and anorectic and developed a mild (one dog) to severe diarrhea five days postinfection. The remaining dogs had subclinical infections but developed a lymphopenia followed by a transient lymphocytosis. The ill dogs developed mild (one dog) to severe neutropenia and a moderate lymphopenia. One died nine days postinfection. Recovery was associated with cessation of viral excretion and with lymphocytosis and antibody production. Two of three dogs challenged intragastrically developed mild clinical signs and a moderate panleukopenia four to eight days postinfection. The pathological changes of the experimental disease were very similar to that of spontaneous disease. Bone marrow changes included a severe granulocytic and mild erythroid depletion. The pathogenesis of canine parvovirus infection is discussed. Images Fig. 4. Fig. 5. Fig. 6. PMID:7340906
Nemeth, Nicole M; Oesterle, Paul T; Poulson, Rebecca L; Jones, Cheryl A; Tompkins, S Mark; Brown, Justin D; Stallknecht, David E
European Starlings (Sturnus vulgaris) and House Sparrows (Passer domesticus) are common peridomestic passerine birds that are often associated with domestic animal production facilities. This association provides a potential means for pathogen transmission between facilities. We inoculated European Starlings and House Sparrows with three non-avian influenza virus strains: two swine isolates (H1N1 and H3N2) and one human isolate representing the H1N1 pandemic strain that originated from swine. No viral shedding was observed in House Sparrows, and shedding was minimal and transient in two of 12 (17%) European Starlings. One of these two infected Starlings seroconverted 14 days after inoculation. These results suggest that these two passerine species are minimally susceptible to current influenza viruses in domestic pigs and therefore pose a negligible risk for transmission between or within swine production facilities.
Wernike, Kerstin; Hoffmann, Bernd; Bréard, Emmanuel; Bøtner, Anette; Ponsart, Claire; Zientara, Stéphan; Lohse, Louise; Pozzi, Nathalie; Viarouge, Cyril; Sarradin, Pierre; Leroux-Barc, Céline; Riou, Mickael; Laloy, Eve; Breithaupt, Angele; Beer, Martin
Since late 2011, a novel orthobunyavirus, named Schmallenberg virus (SBV), has been implicated in many cases of severely malformed bovine and ovine offspring in Europe. In adult cattle, SBV is known to cause a mild transient disease; clinical signs include short febrile episodes, decreased milk production and diarrhoea for a few days. However, the knowledge about clinical signs and pathogenesis in adult sheep is limited. In the present study, adult sheep of European domestic breeds were inoculated with SBV either as cell culture grown virus or as virus with no history of passage in cell cultures. Various experimental set-ups were used. Sampling included blood collection at different time points during the experimental period and selected organ material at autopsy. Data from this study showed, that the RNAemic period in sheep was as short as reported for cattle; viral genome was detectable for about 3-5 days by real-time RT-PCR. In total, 13 out of 30 inoculated sheep became RNAemic, with the highest viral load in animals inoculated with virus from low cell culture passaged or the animal passaged material. Contact animals remained negative throughout the study. One RNAemic sheep showed diarrhoea for several days, but fever was not recorded in any of the animals. Antibodies were first detectable 10-14 days post inoculation. Viral RNA was detectable in spleen and lymph nodes up to day 44 post inoculation. In conclusion, as described for cattle, SBV-infection in adult sheep predominantly results in subclinical infection, transient RNAemia and a specific antibody response. Maintenance of viral RNA in the lymphoreticular system is observed for an extended period.
Villain, Patricia; Gonzalez, Paula; Almonte, Maribel; Franceschi, Silvia; Dillner, Joakim; Anttila, Ahti; Park, Jin Young; De Vuyst, Hugo; Herrero, Rolando
Of the 2,635,000 new cancer cases (excluding non-melanoma skin cancers) occurring in the European Union (EU) in 2012, it is estimated that approximately 185,000 are related to infection with human papillomaviruses (HPVs), hepatitis B and C viruses (HBV and HCV), and Helicobacter pylori (H. pylori). Chronic infection with these agents can lead to cancers of the cervix uteri, liver, and stomach, respectively. Chronic infection with HCV can also lead to B-cell non-Hodgkin lymphoma. Human immunodeficiency virus (HIV) infection continues to be of major public health importance in several EU countries and increases cancer risk via HIV-induced immunosuppression. The fourth edition of the European Code Against Cancer presents recommendations on effective and safe preventive interventions in order to reduce the risk of infection-related cancers in EU citizens. Based on current available evidence, the fourth edition recommends that parents ensure the participation of their children in vaccination programs against HBV (for newborns) and HPV (for girls). In the 'Questions and Answers' (Q&As) section about vaccination and infections in the website for the European Code Against Cancer, individuals who are at risk of chronic HBV or HCV are advised to seek medical advice about testing and obtaining treatment when appropriate. Individuals most at risk of HIV are advised to consult their doctor or healthcare provider to access counselling and, if needed, testing and treatment without delay. Information about H. pylori testing and treatment is also provided as testing might currently be offered in some high-risk areas in Europe. The rationale and supporting evidence for the recommendations on vaccination in the European Code Against Cancer, and for the main recommendations on vaccination and infection in the Q&As, are explained in the present review.
Paredes, Katihuska; Capilla, Javier; Sutton, Deanna A; Mayayo, Emilio; Fothergill, Annette W; Guarro, Josep
We have evaluated the efficacy of amphotericin B, posaconazole, and voriconazole in immunosuppressed murine models of disseminated infection by Curvularia spicifera and Curvularia hawaiiensis. The 3 antifungals improved survival of mice in comparison to controls; however, only the 2 azoles were able to reduce significantly the fungal load.
Vericimo, M A; Figueiredo, I; Teixeira, G A P B; Clemente, S C São
Anisakidosis is a human parasitic disease caused by infections with members of the Anisakidae family. Accidental infection after fish intake affects the gastrointestinal tract as a consequence of mechanical damage caused by migrating larvae. Infections can also trigger allergies, hives, severe asthma or anaphylaxis with angioedema. Although mouse models of intraperitoneal antigenic stimulation exist, enabling immunological studies, few models using gastric introduction of live larvae are available for the study of immunological and gastrointestinal damage in mice. This study was designed to characterize serum reactivity against Anisakis spp. and Contracaecum spp. in Balb/c mice following orogastric inoculation and to assess gastrointestinal damage. These anisakid species were classified at the Universidade Federal Fluminense (UFF) School of Veterinary Medicine and materials for live larval inoculation were developed at the UFF Immunobiology laboratory. Live larvae were inoculated following injection with a NaCl solution. Blood samples were collected and sera screened for immunoglobulin (Ig)E and IgG anti-larva responses to both nematodes, specific for somatic and excretory/secretory antigens, by enzyme-linked immunosorbent assay (ELISA). The means of the optical densities were analysed using analysis of variance (ANOVA) with Tukey's post-hoc test and the general linear model. This analysis identified the presence of anti-IgG seroreactivity to both somatic and excretory/secretory Anisakis antigens in inoculated animals compared with controls (P< 0.001), and no gastric or intestinal damage was observed. These experiments demonstrated that introduction of live Contracaecum spp. into the gastrointestinal tract did not elicit serum sensitization in animals.
Dajani, Adnan S.; Clyde, Wallace A.; Denny, Floyd W.
The pathogenesis of Mycoplasma pneumoniae infection was studied in the Syrian hamster with qualitative and quantitative culture methods and special histopathologic techniques. The animals were readily infected with the mycoplasma, which multiplied throughout the respiratory tract. Sensitivity of this experimental host to infection was indicated by the 50 per cent infective dose, which was 10 colony-forming units of the organism. Inoculation consistently resulted in the production of peribronchial pneumonitis which was induced by the mycoplasma. The organisms were visualized in a superficial location in the mucosa of involved bronchi, by means of indirect fluorescent antibody staining and by a modification of the Brown and Brenn technique. The data indicate applicability of the hamster to the study of problems concerned with M. pneumoniae disease which are impractical or impossible to resolve in the human host. PMID:14319403
de Godoy, Isabela; de Campos, Camila Gonçalves; Pescador, Caroline Argenta; Galceran, João Vitor Amorim; Cândido, Stéfhano Luis; Dutra, Valéria; Nakazato, Luciano
Conidiobolomycosis is an emerging entomophthoramycosis caused by fungi Conidiobolus spp. Animal models are essential for the study of infectious disease in various areas such as pathogenesis, diagnostic methods, treatment and prevention. There is not currently an animal model for conidiobolomycosis. The aim of this study was to create an experimental infection protocol for Conidiobolus lamprauges in gerbils (Meriones unguiculatus). The study animals were randomly divided into four groups of four animals: immunosuppressed with cyclophosphamide (CPA) and infected with C. lamprauges (G1), immunocompetent and infected with C. lamprauges (G2), immunosuppressed with CPA (G3), and an immunocompetent control group (G4). Clinical signs were observed only in G1 animals, where the mortality rate reached 75% by day 7 after infection (AI) with a median survival of 2 days. C. lamprauges was detected only in G1, both by PCR and by isolation. Necropsies of the G1 animals showed lesions in the nasal cavity and lung tissue. These lesions were characterized by polymorphonuclear infiltrate cells and by the presence of hyphal structures under silver staining. This animal model will be useful for further investigation of diseases caused by C. lamprauges, particularly of those associated with immunosuppression factors in naturally occurring animal infections.
Reyes, José L; Espinoza-Jiménez, Arlett F; González, Marisol I; Verdin, Leticia; Terrazas, Luis I
Helminth infections induce strong immunoregulation that can modulate subsequent pathogenic challenges. Taenia crassiceps causes a chronic infection that induces a Th2-biased response and modulates the host cellular immune response, including reduced lymphoproliferation in response to mitogens, impaired antigen presentation and the recruitment of suppressive alternatively activated macrophages (AAMФ). In this study, we aimed to evaluate the ability of T. crassiceps to reduce the severity of experimental autoimmune encephalomyelitis (EAE). Only 50% of T. crassiceps-infected mice displayed EAE symptoms, which were significantly less severe than uninfected mice. This effect was associated with both decreased MOG-specific splenocyte proliferation and IL-17 production and limited leukocyte infiltration into the spinal cord. Infection with T. crassiceps induced an anti-inflammatory cytokine microenvironment, including decreased TNF-α production and high MOG-specific production of IL-4 and IL-10. While the mRNA expression of TNF-α and iNOS was lower in the brain of T. crassiceps-infected mice with EAE, markers for AAMФ were highly expressed. Furthermore, in these mice, there was reduced entry of CD3(+)Foxp3(-) cells into the brain. The T. crassiceps-induced immune regulation decreased EAE severity by dampening T cell activation, proliferation and migration to the CNS.
The European Summer School on Experimental Nuclear Astrophysics has reached the sixth edition, marking the tenth year's anniversary. The spirit of the school is to provide a very important occasion for a deep education of young researchers about the main topics of experimental nuclear astrophysics. Moreover, it should be regarded as a forum for the discussion of the last-decade research activity. Lectures are focused on various aspects of primordial and stellar nucleosynthesis, including novel experimental approaches and detectors, indirect methods and radioactive ion beams. Moreover, in order to give a wide educational offer, some lectures cover complementary subjects of nuclear astrophysics such as gamma ray astronomy, neutron-induced reactions, short-lived radionuclides, weak interaction and cutting-edge facilities used to investigate nuclear reactions of interest for astrophysics. Large room is also given to young researcher oral contributions. Traditionally, particular attention is devoted to the participation of students from less-favoured countries, especially from the southern coast of the Mediterranean Sea. The school is organised by the Catania Nuclear Astrophysics research group with the collaboration of Dipartimento di Fisica e Astromomia - Università di Catania and Laboratori Nazionali del Sud - Istituto Nazionale di Fisica Nucleare.
Freda, Carmela; Funiciello, Francesca; Meredith, Phil; Sagnotti, Leonardo; Scarlato, Piergiorgio; Troll, Valentin R.; Willingshofer, Ernst
Integrating Earth Sciences infrastructures in Europe is the mission of the European Plate Observing System (EPOS).The integration of European analytical, experimental, and analogue laboratories plays a key role in this context and is the task of the EPOS Working Group 6 (WG6). Despite the presence in Europe of high performance infrastructures dedicated to geosciences, there is still limited collaboration in sharing facilities and best practices. The EPOS WG6 aims to overcome this limitation by pushing towards national and trans-national coordination, efficient use of current laboratory infrastructures, and future aggregation of facilities not yet included. This will be attained through the creation of common access and interoperability policies to foster and simplify personnel mobility. The EPOS ambition is to orchestrate European laboratory infrastructures with diverse, complementary tasks and competences into a single, but geographically distributed, infrastructure for rock physics, palaeomagnetism, analytical and experimental petrology and volcanology, and tectonic modeling. The WG6 is presently organizing its thematic core services within the EPOS distributed research infrastructure with the goal of joining the other EPOS communities (geologists, seismologists, volcanologists, etc...) and stakeholders (engineers, risk managers and other geosciences investigators) to: 1) develop tools and services to enhance visitor programs that will mutually benefit visitors and hosts (transnational access); 2) improve support and training activities to make facilities equally accessible to students, young researchers, and experienced users (training and dissemination); 3) collaborate in sharing technological and scientific know-how (transfer of knowledge); 4) optimize interoperability of distributed instrumentation by standardizing data collection, archive, and quality control standards (data preservation and interoperability); 5) implement a unified e-Infrastructure for data
Piranda, Eliane M; Faccini, João Luiz H; Pinter, Adriano; Pacheco, Richard C; Cançado, Paulo H D; Labruna, Marcelo B
We evaluated if Rickettsia rickettsii-experimentally infected dogs could serve as amplifier hosts for hipicephalus sanguineus ticks. In addition, we checked if Rh. sanguineus ticks that acquired Ri. rickettsii from dogs could transmit the bacterium to susceptible hosts (vector competence), and if these ticks could maintain the bacterium by transstadial and transovarial transmissions. Uninfected larvae, nymphs, and adults of Rh. sanguineus were allowed to feed upon three groups of dogs: groups 1 (G1) and 2 (G2) composed of Ri. rickettsii-infected dogs, infected intraperitoneally and via tick bites, respectively, and group 3 composed of uninfected dogs. After larval and nymphal feeding on rickettsemic dogs, 7.1-15.2% and 35.8-37.9% of the molted nymphs and adults, respectively, were shown by polymerase chain reaction (PCR) to be infected by Ri. rickettsii, confirming that both G1 and G2 dogs were efficient sources of rickettsial infection (amplifier host), resulting in transstadial transmission of the agent. These infected nymphs and adults successfully transmitted Ri. rickettsii to guinea pigs, confirming vector competence after acquisition of the infection from rickettsemic dogs. Transovarial transmission of Ri. rickettsii was observed in engorged females that had been infected as nymphs by feeding on both G1 and G2 dogs, but not in engorged females that acquired the infection during adult feeding on these same dogs. In the first case, filial infection rates were generally <50%. No tick exposed to G3 dogs was infected by rickettsiae in this study. No substantial mortality difference was observed between Ri. rickettsii-infected tick groups (G1 and G2) and uninfected tick group (G3). Our results indicate that dogs can be amplifier hosts of Ri. rickettsii for Rh. sanguineus, although only a minority of immature ticks (<45%) should become infected. It appears that Rh. sanguineus, in the absence of horizontal transmission, would not maintain Ri. rickettsii through
Amengual, Blanca; Abellán, Carlos; Bourhy, Hervé
From 1992 to 2000, 976 sera, 27 blood pellets, and 91 brains were obtained from 14 bat species in 37 localities in Spain. Specific anti-European bat lyssavirus 1 (EBL1)-neutralizing antibodies have been detected in Myotis myotis, Miniopterus schreibersii, Tadarida teniotis, and Rhinolophus ferrumequinum in the region of Aragon and the Balearic Islands. Positive results were also obtained by nested reverse transcription-polymerase chain reaction on brain, blood pellet, lung, heart, tongue, and esophagus-larynx-pharynx of M. myotis, Myotis nattereri, R. ferrumequinum, and M. schreibersii. Determination of nucleotide sequence confirmed the presence of EBL1 RNA in the different tissues. In one colony, the prevalence of seropositive bats over time corresponded to an asymmetrical curve, with a sudden initial increase peaking at 60% of the bats, followed by a gradual decline. Banded seropositive bats were recovered during several years, indicating that EBL1 infection in these bats was nonlethal. At least one of this species (M. schreibersii) is migratory and thus could be partially responsible for the dissemination of EBL1 on both shores of the Mediterranean Sea. PMID:11971777
López Sanmartín, Monserrat; Power, Deborah M; de la Herrán, Roberto; Navas, José I; Batista, Frederico M
Ostreid herpesvirus 1 (OsHV-1) infections have been reported in several bivalve species. Mortality of Pacific oyster Crassostrea gigas spat has increased considerably in Europe since 2008 linked to the spread of a variant of OsHV-1 called μvar. In the present study we demonstrated that O. edulis juveniles can be infected by OsHV-1μvar when administered as an intramuscular injection. Mortality in the oysters injected with OsHV-1μvar was first detected 4 days after injection and reached 25% mortality at day 10. Moreover, the high viral load observed and the detection of viral transcripts by in situ hybridization in several tissues of dying oysters suggested that OsHV-1μvar was the cause of mortality in the O. edulis juveniles. This is therefore the first study to provide evidence about the pathogenicity of OsHV-1μvar in a species that does not belong to the Crassostrea genus. Additionally, we present a novel method to detect OsHV-1 transcripts in infected individuals' using in situ hybridization.
Gaze, S; Bethony, J M; Periago, M V
Human hookworm infection is one amongst the most prevalent of the neglected tropical diseases. An informative experimental animal model, that is, one that parallels a human infection, is not available for the study of human hookworm infection. Much of our current understanding of the human immune response during hookworm infection relies on the studies from experimental infection of hookworm-naïve individuals or the natural infections from individuals residing in hookworm-endemic areas. The experimental human infections tend to be acute, dose-controlled infections, often with a low larval inoculum so that they are well tolerated by human volunteers. Natural hookworm infections usually occur in areas where hookworm transmission is constant and infection is chronic. In cases where there has been drug administration in an endemic area, re-infection often occurs quickly even amongst those who were treated. Hence, although many of the characteristics of experimental and natural hookworm infection differ, both models have elements in common: mainly an intense Th2 response with the production of total and specific IgE as well as elevated levels of eosinophilia, IL-5, IL-10 and TNF. While hookworm infection affects millions of individuals worldwide, much of the human immunology of this infection still needs to be studied and understood.
Maccallum, Donna M
Although normally commensals in humans, Candida albicans, Candida tropicalis, Candida parapsilosis, Candida glabrata, and Candida krusei are capable of causing opportunistic infections in individuals with altered physiological and/or immunological responses. These fungal species are linked with a variety of infections, including oral, vaginal, gastrointestinal, and systemic infections, with C. albicans the major cause of infection. To assess the ability of different Candida species and strains to cause infection and disease requires the use of experimental infection models. This paper discusses the mucosal and systemic models of infection available to assay Candida virulence and gives examples of some of the knowledge that has been gained to date from these models.
Poskin, A; Martinelle, L; Mostin, L; Van Campe, W; Dal Pozzo, F; Saegerman, C; Cay, A B; De Regge, N
Schmallenberg virus (SBV) is an orthobunyavirus affecting European domestic ruminants. In this study, the dose-dependent effect of experimental infection of sheep with SBV was evaluated. Four groups of three ewes were each inoculated subcutaneously with 1 mL of successive 10-fold dilutions of an SBV infectious serum. The ewes were monitored for 10 days, but no clinical signs were observed. The number of productively infected animals within each group, as evidenced by viraemia, seroconversion and viral RNA in the organs, depended on the inoculated dose, indicating that a critical dose has to be administered to obtain a homogeneous response in infected animals under experimental conditions. In the productively infected animals, no statistical differences between the different inoculation doses were found in the duration or quantity of viral RNA circulating in blood, nor in the amount of viral RNA present in virus positive lymphoid organs.
This teaching guide focuses on the Southern European food heritage. It is part of the Louisville Area Ethnic Heritage Studies Project described in ED 150 043. The project materials are designed to foster communication across intercultural/ethnic lines. The objective of this unit is to gain familiarity with and appreciate foods from Spain, France,…
This teaching guide focuses on the Northern European food heritage. It is part of the Louisville Area Ethnic Heritage Studies Project described in ED 150 043. The materials are designed to foster communication across intercultural/ethnic lines. The objective of this unit is to gain familiarity with and appreciate foods from Scandinavia, the Soviet…
Song, Young-Jo; Park, Woo-Jung; Park, Byung-Joo; Kwak, Sang-Woo; Kim, Yong-Hyeon; Lee, Joong-Bok; Park, Seung-Yong; Song, Chang-Seon; Lee, Sang-Won; Seo, Kun-Ho; Kang, Young-Sun; Park, Choi-Kyu; Song, Jae-Young; Choi, In-Soo
Hepatitis A virus (HAV) is the leading cause of acute viral hepatitis worldwide, with HAV infection being restricted to humans and nonhuman primates. In this study, HAV infection status was serologically determined in domestic pigs and experimental infections of HAV were attempted to verify HAV infectivity in pigs. Antibodies specific to HAV or HAV-like agents were detected in 3.5% of serum samples collected from pigs in swine farms. When the pigs were infected intravenously with 2 × 10(5) 50% tissue culture infectious dose (TCID50 ) of HAV, shedding of the virus in feces, viremia, and seroconversion were detected. In pigs orally infected with the same quantity of HAV, viral shedding was detected only in feces. HAV genomic RNA was detected in the liver and bile of intravenously infected pigs, but only in the bile of orally infected pigs. In further experiments, pigs were intravenously infected with 6 × 10(5) TCID50 of HAV. Shedding of HAV in feces, along with viremia and seroconversion, were confirmed in infected pigs but not in sentinel pigs. HAV genomic RNA was detected in the liver, bile, spleen, lymph node, and kidney of the infected pigs. HAV antigenomic RNA was detected in the spleen of one HAV-infected pig, suggesting HAV replication in splenic cells. Infiltration of inflammatory cells was observed in the livers of infected pigs but not in controls. This is the first experimental evidence to demonstrate that human HAV strains can infect pigs.
Souto, S; Lopez-Jimena, B; Alonso, M C; García-Rosado, E; Bandín, I
The susceptibility of juvenile European sea bass and Senegalese sole to three VNNV isolates (a reassortant RGNNV/SJNNV, as well as the parental RGNNV and SJNNV genotypes) has been evaluated by challenges using two inoculation ways (bath and intramuscular injection). The results demonstrate that these two fish species are susceptible to all the VNNV isolates tested. In European sea bass, RGNNV caused the highest cumulative mortality, reaching maximum values of viral RNA and titres. Although the SJNNV isolate did not provoke mortality or clinical signs of disease in this fish species, viral production in survivor fish was determined; on the other hand the reassortant isolate did cause mortality and clinical signs of disease, although less evident than those recorded after RGNNV infection. These results suggest that the changes suffered by the SJNNV RNA2 segment of the reassortant isolate, compared to the parental SJNNV, may have involved host-specificity and/or virulence determinants for European sea bass. Regarding Senegalese sole, although the three isolates caused 100% mortality, the reassortant strain provoked the most acute symptoms, and more quickly, especially in the bath challenge. This was also the isolate showing less difference between the number of RNA copies and viral titre, reaching the highest titres of infective viral particles in nervous tissue of infected animals. The RGNNV isolate produced the lowest values of infective viral particles. All these results suggest that the RGNNV and the reassortant isolates are the most suited for infecting European sea bass and Senegalese sole, respectively.
Tomlinson, A J; Chambers, M A; Carter, S P; Wilson, G J; Smith, G C; McDonald, R A; Delahay, R J
The behaviour of certain infected individuals within socially structured populations can have a disproportionately large effect on the spatio-temporal distribution of infection. Endemic infection with Mycobacterium bovis in European badgers (Meles meles) in Great Britain and Ireland is an important source of bovine tuberculosis in cattle. Here we quantify the risk of infection in badger cubs in a high-density wild badger population, in relation to the infection status of resident adults. Over a 24-year period, we observed variation in the risk of cub infection, with those born into groups with resident infectious breeding females being over four times as likely to be detected excreting M. bovis than cubs from groups where there was no evidence of infection in adults. We discuss how our findings relate to the persistence of infection at both social group and population level, and the potential implications for disease control strategies.
Leang-Chung, Choh; Vellasamy, Kumutha Malar; Mariappan, Vanitha; Li-Yen, Chang; Vadivelu, Jamuna
Burkholderia pseudomallei is an intracellular Gram-negative bacterial pathogen intrinsically resistant to a variety of antibiotics. Phages have been developed for use as an alternative treatment therapy, particularly for bacterial infections that do not respond to conventional antibiotics. In this study, we investigated the use of phages to treat cells infected with B. pseudomallei. Phage C34 isolated from seawater was purified and characterised on the basis of its host range and morphology using transmission electron microscopy (TEM). Phage C34 was able to lyse 39.5% of B. pseudomallei clinical strains. Due to the presence of contractile tail, phage C34 is classified as a member of the family Myoviridae, a tailed double-stranded DNA virus. When 2 × 105 A549 cells were exposed to 2 × 107 PFU of phage C34, 24 hours prior to infection with 2 × 106 CFU of B. pseudomallei, it was found that the survivability of the cells increased to 41.6 ± 6.8% as compared to 22.8 ± 6.0% in untreated control. Additionally, application of phage successfully rescued 33.3% of mice infected with B. pseudomallei and significantly reduced the bacterial load in the spleen of the phage-treated mice. These findings indicate that phage can be a potential antimicrobial agent for B. pseudomallei infections. PMID:27387381
Moazed, T. C.; Kuo, C.; Patton, D. L.; Grayston, J. T.; Campbell, L. A.
Chlamydia pneumoniae (TWAR), a common cause of acute respiratory disease in humans, has recently been associated with coronary and aortic atherosclerosis. In this study, we evaluated rabbit models of chlamydial infection to investigate the pathogenesis of C. pneumoniae infection. New Zealand White rabbits were inoculated intranasally and intratracheally with C. pneumoniae, strain AR-39, and primary and repeated infection were assessed. After a single inoculation, lung pathology was characterized by a moderate self-resolving interstitial pneumonia with bronchiolitis of 21 days in duration. Chlamydial DNA was detected by polymerase chain reaction (PCR) intermittently in the upper respiratory tract and lung tissue through day 21 postinoculation, spleen tissue at day 14, and peripheral blood mononuclear cells at days 3 and 21. After repeated inoculations, chlamydial DNA was detected by PCR in the upper respiratory tract and lung tissue through day 42. Lung lesions consisted of multifocal interstitial mononuclear cell aggregates that persisted up to day 42. Watanabe heritable hyperlipidemic rabbits were less susceptible to C. pneumoniae infection. After multiple inoculations of Watanabe rabbits, C. pneumoniae was detected by PCR and/or immunocytochemistry until day 21. In conclusion, C. pneumoniae induced a moderate respiratory infection in these rabbit models. Images Figure 1 Figure 2 Figure 3 PMID:8579129
Jaton, Laure; Kritikos, Antonios; Bodenmann, Patrick; Greub, Gilbert; Merz, Laurent
Current conflicts in some regions of the world give rise to massive immigration waves. Consequently, some infections that had nearly disappeared in Europe nowadays re-emerge. They are related to the epidemiology of the refugees' origin, but also to the epidemiology of the country crossed during migration. Hygiene conditions, often precarious during the journey, favor their transmission. Thus, cases of louse borne relapsing fever and diphtheria emerge in Europe and in Switzerland since 2074 whereas cutaneous Panton-Valen tine Staphylococcus aureus infection are more commonly observed nowadays.
Ogrzewalska, Maria; Soares, João F.; Martins, Thiago F.; Soares, Herbert S.; Moraes-Filho, Jonas; Nieri-Bastos, Fernanda A.; Almeida, Aliny P.; Pinter, Adriano
We experimentally infected Amblyomma aureolatum ticks with the bacterium Rickettsia rickettsii, the etiologic agent of Rocky Mountain spotted fever (RMSF). These ticks are a vector for RMSF in Brazil. R. rickettsii was efficiently conserved by both transstadial maintenance and vertical (transovarial) transmission to 100% of the ticks through 4 laboratory generations. However, lower reproductive performance and survival of infected females was attributed to R. rickettsii infection. Therefore, because of the high susceptibility of A. aureolatum ticks to R. rickettsii infection, the deleterious effect that the bacterium causes in these ticks may contribute to the low infection rates (<1%) usually reported among field populations of A. aureolatum ticks in RMSF-endemic areas of Brazil. Because the number of infected ticks would gradually decrease after each generation, it seems unlikely that A. aureolatum ticks could sustain R. rickettsii infection over multiple successive generations solely by vertical transmission. PMID:21529391
Labruna, Marcelo B; Ogrzewalska, Maria; Soares, João F; Martins, Thiago F; Soares, Herbert S; Moraes-Filho, Jonas; Nieri-Bastos, Fernanda A; Almeida, Aliny P; Pinter, Adriano
We experimentally infected Amblyomma aureolatum ticks with the bacterium Rickettsia rickettsii, the etiologic agent of Rocky Mountain spotted fever (RMSF). These ticks are a vector for RMSF in Brazil. R. rickettsii was efficiently conserved by both transstadial maintenance and vertical (transovarial) transmission to 100% of the ticks through 4 laboratory generations. However, lower reproductive performance and survival of infected females was attributed to R. rickettsii infection. Therefore, because of the high susceptibility of A. aureolatum ticks to R. rickettsii infection, the deleterious effect that the bacterium causes in these ticks may contribute to the low infection rates (<1%) usually reported among field populations of A. aureolatum ticks in RMSF-endemic areas of Brazil. Because the number of infected ticks would gradually decrease after each generation, it seems unlikely that A. aureolatum ticks could sustain R. rickettsii infection over multiple successive generations solely by vertical transmission.
Corner, L A L; Murphy, D; Costello, E; Gormley, E
The eradication of tuberculosis (Mycobacterium bovis infection) from cattle herds may be compromised if infected wildlife species, such as European badgers (Meles meles), share the same environment and contribute to transfer of infection. Options for dealing with tuberculosis in this wild reservoir host are limited by conservation and social concerns, despite a clear implication that infected badgers are involved with the initiation of tuberculosis in cattle herds. Vaccination of badgers against M. bovis, if successfully employed, would directly facilitate the completion of bovine tuberculosis eradication in affected areas. Vaccine trials in captive badgers have established that the M. bovis bacille Calmette-Guérin (BCG) vaccine can induce a protective response that limits the distribution and severity of tuberculosis disease following experimental challenge. The protective effect of the vaccine has been demonstrated when the vaccine was delivered by subcutaneous injection, deposited on mucous membranes, and given orally in a lipid formulation. A large-scale field trial of oral BCG vaccine has been designed to measure the protection generated in wild badgers subjected to natural transmission of infection and to estimate vaccine efficacy. These parameters will be estimated by comparing the prevalence of M. bovis infection in vaccinated and nonvaccinated badgers. The results will provide a framework for the development and implementation of a national strategy to eliminate the disease in badger populations and if successful will remove this major impediment to bovine tuberculosis eradication.
Beckmann, Katie M; Harris, Eileen; Pocknell, Ann M; John, Shinto K; Macgregor, Shaheed K; Cunningham, Andrew A; Lawson, Becki
A European Robin (Erithacus rubecula) found dead in England had marked blepharitis and periocular alopecia associated with Aprocta cylindrica (Nematoda: Aproctidae) and concurrent mixed fungal infections. Aprocta cylindrica should be considered a differential diagnosis in periocular abnormalities of robins and other insectivorous, migratory passerines in Western Europe.
de Souza, William Marciel; Machado, Alex Martins; Figueiredo, Luiz Tadeu Moraes
This study shows an experimental spillover infection ofSigmodontinae rodents with Rio Mamore hantavirus (RIOMV).Necromys lasiurus and Akodon sp were infected with 103 RNA copies of RIOMV by intraperitoneal administration. The viral genome was detected in heart, lung, and kidney tissues 18 days after infection (ai), and viral excretion in urine and faeces began at four and six ai, respectively. These results reveal that urine and faeces of infected rodents contain the virus for at least 18 days. It is possible that inhaled aerosols of these excreta could transmit hantavirus to humans and other animals. PMID:27223653
Gerke, John R.; Magliocco, Michael V.
Pseudomonas aeruginosa infection of human cornea is rare but serious. The work of previous investigators using experimental infection primarily of rabbit cornea resulted in successful therapy for 10 to 50% of clinical cases. The advantage of using the mouse is demonstrated. The methods we adapted for characterizing the untreated experimental infection included: incising the cornea to enable establishing the infection; corneal examination with a steroscopic microscope; grading corneal pathology; qualitative and quantitative monitoring of the infecting bacteria by culturing and staining sectioned and dissected tissues. The characteristics of the tissue pathology, host response, and infection were similar to those reported for other animals and man. Corneal pathology was frequently nearly maximal 1 day after infection; host response involved a progression of events of long duration; pathology persisted well beyond the period of bacterial infection. The infection was essentially noncommunicable, and invasiveness was limited to the tissues of the incised eye. The results show the possibility of tests for invasiveness of clinical isolates and for screening for therapeutic and prophylactic measures. PMID:16557955
Hudson, Michelle; Balls, Michael
The latest report on experimental animal use in the EU indicates a slight decrease. However, the figures, which are for 2008, show that around 12 million animals are still used in laboratories throughout Europe. To date, only a summary of the data has been published, and so this report is limited to giving a comparison of trends and significant changes. The impact on the figures of the revision of Directive 86/609/EEC and the 7th Amendment to the Cosmetics Directive 76/768/EEC is discussed. In addition, consideration is given to which areas of research and regulation will need to be closely monitored, and the importance of continuing to push for the implementation of Three Rs initiatives is highlighted.
Oliver, Kerry J; Reddin, Karen M; Bracegirdle, Philippa; Hudson, Michael J; Borrow, Ray; Feavers, Ian M; Robinson, Andrew; Cartwright, Keith; Gorringe, Andrew R
Immunological and epidemiological evidence suggests that the development of natural immunity to meningococcal disease results from colonization of the nasopharynx by commensal Neisseria spp., particularly with N. lactamica. We report here that immunization with N. lactamica killed whole cells, outer membrane vesicles, or outer membrane protein (OMP) pools and protected mice against lethal challenge by a number of diverse serogroup B and C meningococcal isolates in a model of bacteremic infection. Sera raised to N. lactamica killed whole cells, OMPs, or protein pools were found to cross-react with meningococcal isolates of a diverse range of genotypes and phenotypes. The results confirm the potential of N. lactamica to form the basis of a vaccine against meningococcal disease.
Newbold, L R; Hockley, F A; Williams, C F; Cable, J; Reading, A J; Auchterlonie, N; Kemp, P S
The effect of Anguillicola crassus, Pseudodactylogyrus bini and Pseudodactylogyrus anguillae infection on the behaviour of downstream migrating adult European eels Anguilla anguilla as they encountered accelerating water velocity, common at engineered structures where flow is constricted (e.g. weirs and bypass systems), was evaluated in an experimental flume. The probability of reacting to, and rejecting, the velocity gradient was positively related to A. crassus larval, adult and total abundance. High abundance of Pseudodactylogyrus spp. reduced this effect, but A. crassus was the strongest parasitic factor associated with fish behaviour, and abundance was positively related to delay in downstream passage. Delayed downstream migration at hydraulic gradients associated with riverine anthropogenic structures could result in additional energetic expenditure for migrating A. anguilla already challenged by A. crassus infection.
Atkins, C E; Arther, R G; Ciszewski, D K; Davis, W L; Ensley, S M; Guity, P S; Chopade, H; Hoss, H; Settje, T L
The safety of heartworm preventives in heartworm-positive cats has traditionally been evaluated using adult Dirofilaria immitis removed from infected dogs and surgically implanted into the cats. An alternate study model uses infective larvae to establish adult infections in cats. Unfortunately, the number of adult worms resulting from the latter method varies widely from none to more than 30, both unacceptable for studies of natural heartworm infection and for studies evaluating product safety in heartworm-infected cats. We sought to determine infection severity in experimental infections via echocardiography to reduce the chances of enrolling uninfected and heavily infected cats into a study. Eighty adult cats were each inoculated with 60 infective D. immitis larvae and maintained for 8 months to allow for the development of adult worms. Antigen and antibody testing, as well as echocardiographic imaging, were performed to confirm and estimate adult worm burdens. Approximately 8 and 12 months post-infection, echocardiographic examination was performed to confirm and enumerate adult D. immitis populations in the cardiovascular system. Worm burdens were stratified as 0, 1-3, 4-11, and > 11 adults, with 0 being considered uninfected and more than 11 considered too heavily infected to be relevant for anthelmintic studies. Cats with clinically relevant infections (1-10 adults) subsequently received multiple treatments with the investigational drug, and worm burdens were confirmed by necropsy 30 days following the final treatment. Worm burden estimated with echocardiography correlated well, but not precisely, with post-mortem counts (p < 0.001, r2 = 0.67). Echocardiography under-, over-, and exactly estimated heartworm burden 53%, 27%, and 22% of the time, respectively. Although the correct category (0-4) was determined by echocardiography in only 54% of cats, positive cats were distinguished from negative cats 88% of the time and the heaviest infections (> 11) were
Gunawardana, Natasha; Finney, Lydia; Johnston, Sebastian L; Mallia, Patrick
Chronic obstructive pulmonary disease (COPD) is a major public health problem and will be one of the leading global causes of mortality over the coming decades. Much of the morbidity, mortality and health care costs of COPD are attributable to acute exacerbations, the commonest causes of which are respiratory infections. Respiratory viruses are frequently detected in COPD exacerbations but direct proof of a causative relationship has been lacking. We have developed a model of COPD exacerbation using experimental rhinovirus infection in COPD patients and this has established a causative relationship between virus infection and exacerbations. In addition it has determined some of the molecular mechanisms linking virus infections to COPD exacerbations and identified potential new therapeutic targets. This new data should stimulate research into the role of antiviral agents as potential treatments for COPD exacerbations. Testing of antiviral agents has been hampered by the lack of a small animal model for rhinovirus infection and experimental rhinovirus infection in healthy volunteers has been used to test treatments for the common cold. Experimental rhinovirus infection in COPD subjects offers the prospect of a model that can be used to evaluate the effects of new treatments for virus-induced COPD exacerbations, and provide essential data that can be used in making decisions regarding large scale clinical trials.
Seymour, C; Kramer, L D; Peralta, P H
Experimental infection of 11 Bradypus variegatus and Choloepus hoffmanni sloths with St. Louis encephalitis (SLE) virus produced detectable viremias of seven to 27 (median 13) days duration and maximum titers of 2.7 to 6.5 (median 5.1) log10 median suckling mouse intracranial lethal doses (SMicLD50) per ml. Experimental SLE viremia onset was delayed and maximum titer depressed in two sloths concurrently infected with naturally acquired viruses. SLE viremias in four experimentally inoculated cormorants Phalacrocorax olivaceus were shorter, and of equal or lower titer, than in sloths. Colonized Culex pipiens quinquefasciatus mosquitoes were infected by feeding on sloths circulating at least 4.8 log10 SMicLD50 of SLE virus per ml, and subsequently transmitted the infection to mice and chicks. An uninoculated baby Bradypus became infected by contact transmission from its mother. The antibody response of sloths to SLE virus was slow, being undetectable until several weeks post-inoculation. However, both sloth species developed high and long-lasting neutralizing and hemagglutination-inhibition antibody titers. The complement-fixation antibody response in Bradypus was lower and slower to develop than in Choloepus. Sloths with naturally acquired SLE virus antibody did not become detectably viremic after experimental inoculation. Neither sloths nor cormorants become overly ill from SLE virus infection.
Freda, C.; Funiciello, F.; Meredith, P.; Sagnotti, L.; Scarlato, P.; Troll, V. R.; Willingshofer, E.; EPOS-WG6
The EU policy for scientific research in the third millennium is that of a coordinated approach to support and develop continent-scale research infrastructures. The vision is to integrate the existing research infrastructures in order to increase the accessibility and usability of multidisciplinary data, enhancing worldwide interoperability by establishing a leading integrated European infrastructure and services. Integrating Earth Sciences infrastructures in Europe is the mission of the European Plate Observing System (EPOS), a research infrastructure and e-science for data and observatories on earthquakes, volcanoes, surface dynamics and tectonics. Within the existing core elements to be integrated in the EPOS cyber-infrastructure are: geographical distributed observational infrastructures (seismic and geodetic networks), observatories for multidisciplinary local data acquisition (e.g., volcanoes, active fault-zone, geothermal and deep drilling experiments), and analytical facilities for data repositories and integration. The integration of European analytical, experimental, and analogue laboratories plays a key role in this context and is the task of EPOS Working Group 6 (WG6). The Analytical and Experimental LaboratoriesGroup thus aims to link experimental, analytical, and analogue laboratories into a single, but geographically distributed, infrastructure for rock physics, including palaeomagnetism, analytical and experimental petrology and volcanology, and tectonic modeling.The WG6 has set a short term goal that has now been achieved, being a review of operational laboratory facilities in the community and the creation of a database from that information. Currently 12 countries (Germany, Greece, Ireland, Italy, Portugal, Romania, Slovenia, Spain, Sweden, Switzerland, The Netherlands, United Kingdom) are included in the database. As long-term goals, the WG6 aims to create mechanisms and procedures for easy access to laboratory facilities, turning small
Michel, Anita L
Mycobacterium fortuitum and at least 1 unidentified species of soil mycobacteria were isolated from lymph nodes from 4 of 5 African buffalo (Syncerus caffer) that had been culled because of positive test results using the Bovigam assay. The buffalo were part of a group of 16 free-ranging buffalo captured in the far north of the Kruger National Park (South Africa) assumed to be free of bovine tuberculosis. No Mycobacterium bovis was isolated. To investigate the possible cause of the apparent false-positive diagnosis, the Mycobacterium isolates were inoculated into 4 experimental cattle and their immune responses monitored over a 13-week period, using the gamma interferon assay. The immune reactivity was predominantly directed toward avian tuberculin purified protein derivative (PPD) and lasted for approximately 8 weeks. During that period 3 of 4 cattle yielded positive test results on 1 or 2 occasions. The immune responsiveness was boosted when the inoculations were repeated after 15 weeks, which led to 2 subsequent positive reactions in the experimental animal that did not react previously. Including an additional stimulatory antigen, sensitin prepared from M. fortuitum in the gamma interferon assay, showed that it was able to elicit a detectable gamma interferon response in all 4 experimentally inoculated cattle when applied in parallel with bovine and avian tuberculin PPD for the stimulation of blood samples. The implications of occasional cross-reactive responses in natural cases of infection with environmental mycobacteria in the diagnosis of bovine tuberculosis in African buffalo and cattle in South Africa are discussed.
Pérez-Ramírez, Elisa; Llorente, Francisco; Jiménez-Clavero, Miguel Ángel
Avian models of West Nile virus (WNV) disease have become pivotal in the study of infection pathogenesis and transmission, despite the intrinsic constraints that represents this type of experimental research that needs to be conducted in biosecurity level 3 (BSL3) facilities. This review summarizes the main achievements of WNV experimental research carried out in wild birds, highlighting advantages and limitations of this model. Viral and host factors that determine the infection outcome are analyzed in detail, as well as recent discoveries about avian immunity, viral transmission, and persistence achieved through experimental research. Studies of laboratory infections in the natural host will help to understand variations in susceptibility and reservoir competence among bird species, as well as in the epidemiological patterns found in different affected areas. PMID:24531334
There is considerable confusion over the management of Helicobacter pylori infection, particularly among primary care physicians, and numerous European countries lack national guidelines in this rapidly growing area of medicine. The European Helicobacter Pylori Study Group therefore organised a meeting in Maastricht of H pylori experts, primary care physicians and representatives of National Societies of Gastroenterology from Europe to establish consensus guidelines on the management of H pylori at the primary care and specialist levels, and to consider general health care issues associated with the infection. As in previous guidelines, eradication therapy was recommended in all H pylori positive patients with peptic ulcer disease. Additionally, at the primary care level in dyspeptic patients < 45 years old and with no alarm symptoms, diagnosis is recommended by non-invasive means (13C urea breath test, serology) and if H pylori positive the patient should be treated. Moreover, at the specialist level the indications for eradication of H pylori were also broadened to include H pylori positive patients with functional dyspepsia in whom no other possible causes of symptoms are identified by the specialist (after a full investigation including endoscopy, ultrasound and other necessary investigations), patients with low grade gastric mucosa associated lymphoid tissue (MALT) lymphoma (managed in specialised centres) and those with gastritis with severe macro- or microscopic abnormalities. There was consensus that treatment regimens should be simple, well tolerated and achieve an eradication rate of over 80% on an intention to treat basis. It was strongly recommended, therefore, that eradication treatment should be with proton pump inhibitor based triple therapy for seven days, using a proton pump inhibitor and two of the following: clarithromycin, a nitroimidazole (metronidazole or tinidazole) and amoxycillin. PMID:9274464
Huffman, J E; Roscoe, D E
Sphaeridiotrema globulus in experimentally infected mute swans (Cygnus olor), mallard ducks (Anas platyrhynchos) and Canada geese (Branta canadensis) induced ulcerative hemorrhagic enteritis. Sites of infection include the jejunum and ileum. The digeneans ulcerated the intestine. The inflammatory response was primarily lymphocytic with some eosinophils. Severe hemorrhage from damaged submucosal capillaries provided a blood meal for the parasite and caused anemia in the host. Extra-medullary hematopoiesis occurred in the liver, and an erythroid hyperplasia occurred in the bone marrow of infected birds. Infected birds exhibited muscular weakness and died from shock associated with severe blood loss. Mallards and Canada geese were less susceptible to fatal infection than the mute swan as evidenced by survivors in the higher dose groups.
Onunkwo, Charles C; Hahn, Beth L; Sohnle, Peter G
Staphylococcal skin infections are quite common in human patients. These infections often clear spontaneously, but may also progress locally and/or disseminate to cause serious and sometimes fatal deep infections. The present studies were undertaken to examine the clearance phase of experimental cutaneous Staphylococcus aureus infections in a mouse model system. Previous work in this system has shown that staphylococci applied to the skin rapidly disseminate to the spleen and kidney. In the present experiments the bacteria were found to persist at the skin infection site at a time (8 days after inoculation) when they had disappeared from the spleen and kidney. Examination of the infected skin at earlier times revealed rapid (within 6 h) invasion into the stratum corneum, stratum Malpighii, and dermis, but subsequent redistribution of bacteria (at 1-2 days) to more superficial sites, particularly crusts located just above the skin surface. The crusts seen in these infections were of two distinct types, which were termed type 1 and type 2. Type 1 crusts appeared first, consisted of bacteria, inflammatory cells, and debris, and developed over an intact epidermis. Type 2 crusts arose from the process of dermal necrosis previously reported to take place at 2 days in this model system. In the latter situation the bacteria were not really cleared from the epidermis and dermis; rather those layers were transformed into a superficial crust that contained the bacteria. Deep hair follicle infections in the dermis were found in these infections, but they did not persist and did not seem to be a reservoir for organisms in the dermis. Resolution of these experimental infections appeared to involve redistribution of invading bacteria to more superficial locations in crusts above the skin surface, marked proliferation of the epidermis, loss of the bacteria-laden crusts from the skin, and eventual healing of the cutaneous damage.
Hendra (HeV) and Nipah (NiV) viruses (genus Henipavirus (HNV; family Paramyxoviridae) are emerging zoonotic agents that can cause severe respiratory distress and acute encephalitis in humans. Given the lack of effective therapeutics and vaccines for human use, these viruses are considered as public health concerns. Several experimental animal models of HNV infection have been developed in recent years. Here, we review the current status of four of the most promising experimental animal models (mice, hamsters, ferrets, and African green monkeys) and their suitability for modeling the clinical disease, transmission, pathogenesis, prevention, and treatment for HNV infection in humans.
Hall, Jeffrey S.; Ip, Hon S.; Teslaa, Joshua L.; Nashold, Sean W.; Dusek, Robert
In 2013 a novel avian influenza H7N9 virus was isolated from several critically ill patients in China, and infection with this virus has since caused more than 200 human deaths. Live poultry markets are the likely locations of virus exposure to humans. Peridomestic avian species also may play important roles in the transmission and maintenance of H7N9 at live poultry markets. We experimentally challenged wild European Starlings (Sturnus vulgaris) with the novel H7N9 virus and measured virus excretion, clinical signs, and infectious dose. We found that European Starlings can be infected with this virus when inoculated with relatively high doses, and we predict that infected birds excrete sufficient amounts of virus to transmit to other birds, including domestic chickens. Infected European Starlings showed no clinical signs or mortality after infection with H7N9. This abundant peridomestic bird may be a source of the novel H7N9 virus in live poultry markets and may have roles in virus transmission to poultry and humans.
Jackson, A C; Phelan, C C; Rossiter, J P
Rabies virus is a highly neuronotropic virus and glial cell infection is not prominent in the central nervous system (CNS). Paraffin-embedded tissues from the cerebella of skunks experimentally infected with either a skunk salivary gland isolate of street rabies virus or the challenge virus standard (CVS) strain of fixed rabies virus were examined with immunoperoxidase staining for rabies virus antigen by using an anti-rabies virus nucleocapsid protein monoclonal antibody. A skunk infected with street rabies virus showed prominent infection of Bergmann glia. Although infected Purkinje cells were observed, they usually demonstrated a relatively small amount of antigen in their perikarya. A CVS-infected skunk showed many intensely labeled Purkinje cells and a relatively small number of infected Bergmann glia. These findings indicate that although rabies virus is a highly neuronotropic virus, street rabies virus strains do not always demonstrate strict neuronotropism in the central nervous system. Images Figure 1. PMID:11041500
Jackson, A C; Phelan, C C; Rossiter, J P
Rabies virus is a highly neuronotropic virus and glial cell infection is not prominent in the central nervous system (CNS). Paraffin-embedded tissues from the cerebella of skunks experimentally infected with either a skunk salivary gland isolate of street rabies virus or the challenge virus standard (CVS) strain of fixed rabies virus were examined with immunoperoxidase staining for rabies virus antigen by using an anti-rabies virus nucleocapsid protein monoclonal antibody. A skunk infected with street rabies virus showed prominent infection of Bergmann glia. Although infected Purkinje cells were observed, they usually demonstrated a relatively small amount of antigen in their perikarya. A CVS-infected skunk showed many intensely labeled Purkinje cells and a relatively small number of infected Bergmann glia. These findings indicate that although rabies virus is a highly neuronotropic virus, street rabies virus strains do not always demonstrate strict neuronotropism in the central nervous system.
Gibert, Patricia; Martinez, Julien; Fraimout, Antoine; Jiggins, Francis; Andrieux, Thibault; Siozios, Stefanos; Anfora, Gianfranco; Miller, Wolfgang; Rota-Stabelli, Omar; Mouton, Laurence
The invasive pest Drosophila suzukii is characterized by a specific fresh-fruit targeting behavior and has quickly become a menace for the fruit economy of newly infested North American and European regions. D. suzukii carries a strain of the endosymbiotic bacterium Wolbachia, named wSuz, which has a low infection frequency and no reproductive manipulation capabilities in American populations of D. suzukii. To further understand the nature of wSuz biology and assess its utility as a tool for controlling this pest’s populations, we investigated the prevalence of Wolbachia in 23 European D. suzukii populations, and compared our results with those available in American populations. Our data showed a highly variable infection frequency with a mean prevalence of 46%, which is significantly higher than the 17% found in American populations. Based on Multilocus Sequence Typing analysis, a single wSuz strain was diagnosed in all European populations of D. suzukii. In agreement with American data, we found no evidence of cytoplasmic incompatibility induced by wSuz. These findings raise two questions: a) why Wolbachia is maintained in field populations of D. suzukii and b) what are the selective forces responsible for the variation in prevalence within populations, particularly between European and American continents? Our results provide new insights into the D. suzukii-Wolbachia association and highlight regional variations that await further investigation and that should be taken into account for using Wolbachia-based pest management programs. PMID:26809119
Cattel, Julien; Kaur, Rupinder; Gibert, Patricia; Martinez, Julien; Fraimout, Antoine; Jiggins, Francis; Andrieux, Thibault; Siozios, Stefanos; Anfora, Gianfranco; Miller, Wolfgang; Rota-Stabelli, Omar; Mouton, Laurence
The invasive pest Drosophila suzukii is characterized by a specific fresh-fruit targeting behavior and has quickly become a menace for the fruit economy of newly infested North American and European regions. D. suzukii carries a strain of the endosymbiotic bacterium Wolbachia, named wSuz, which has a low infection frequency and no reproductive manipulation capabilities in American populations of D. suzukii. To further understand the nature of wSuz biology and assess its utility as a tool for controlling this pest's populations, we investigated the prevalence of Wolbachia in 23 European D. suzukii populations, and compared our results with those available in American populations. Our data showed a highly variable infection frequency with a mean prevalence of 46%, which is significantly higher than the 17% found in American populations. Based on Multilocus Sequence Typing analysis, a single wSuz strain was diagnosed in all European populations of D. suzukii. In agreement with American data, we found no evidence of cytoplasmic incompatibility induced by wSuz. These findings raise two questions: a) why Wolbachia is maintained in field populations of D. suzukii and b) what are the selective forces responsible for the variation in prevalence within populations, particularly between European and American continents? Our results provide new insights into the D. suzukii-Wolbachia association and highlight regional variations that await further investigation and that should be taken into account for using Wolbachia-based pest management programs.
Rivera-Benitez, José Francisco; Martínez-Bautista, Rebeca; Pérez-Torres, Armando; García-Contreras, Adelfa Del Carmen; Reyes-Leyva, Julio; Hernández, Jesús; Ramírez-Mendoza, Humberto
Porcine rubulavirus is the etiological agent of blue eye disease in pigs. In boars, this virus causes orchitis and epididymitis and reduces seminal quality. The objective of this study was to determine the persistence of porcine rubulavirus in experimentally infected boars. Nine 12-month-old boars were infected with 5 ml of the PAC-3 strain of porcine rubulavirus at 1 × 10(5) TCID(50)/ml and held for 142 days post infection (DPI) to evaluate humoral immune response. The virus was isolated in cell cultures and detected by RT-PCR. Infection with porcine rubulavirus produced clinical signs beginning at 5 DPI. Necropsy results showed that 3 boars had lesions in the testicles and epididymes. Histological analysis showed the characteristic lesions in all infected boars. Porcine rubulavirus antibodies were detected in the second week post infection and increased significantly (P<0.05) over time. Isolation of the virus from semen was achieved between 5 DPI and 48 DPI and from the testicles and epididymes between 64 DPI and 142 DPI. Viral RNA was detected in the serum between 2 DPI and 64 DPI and in the semen until 142 DPI. These results confirm that the RNA of the porcine rubulavirus persists in the semen and that this virus remains in the reproductive tract for prolonged periods of infection. Semen of persistently infected boars, therefore, represents an important source of the virus and a risk factor for the spread of blue eye disease in swine populations.
Hamoen, Marleen; Broekhuizen, Berna DL; Little, Paul; Melbye, Hasse; Coenen, Samuel; Goossens, Herman; Butler, Chris C; Francis, Nick A; Verheij, Theo JM
Background It is largely unknown what medication is used by patients with lower respiratory tract infection (LRTI). Aim To describe the use of self-medication and prescribed medication in adults presenting with LRTI in different European countries, and to relate self-medication to patient characteristics. Design and setting An observational study in 16 primary care networks in 12 European countries. Method A total of 2530 adult patients presenting with LRTI in 12 European countries filled in a diary on any medication used before and after a primary care consultation. Patient characteristics related to self-medication were determined by univariable and multivariable logistic regression analysis. Results The frequency and types of medication used differed greatly between European countries. Overall, 55.4% self-medicated before consultation, and 21.5% after consultation, most frequently with paracetamol, antitussives, and mucolytics. Females, non-smokers, and patients with more severe symptoms used more self-medication. Patients who were not prescribed medication during the consultation self-medicated more often afterwards. Self-medication with antibiotics was relatively rare. Conclusion A considerable amount of medication, often with no proven efficacy, was used by adults presenting with LRTI in primary care. There were large differences between European countries. These findings should help develop patient information resources, international guidelines, and international legislation concerning the availability of over-the-counter medication, and can also support interventions against unwarranted variations in care. In addition, further research on the effects of symptomatic medication is needed. PMID:24567621
Gwaltney, J M; Hendley, J O
Transfer of experimental rhinovirus infection by an intermediary environmental surface was examined in healthy young adults, in four studies done in 1980--1981, by having recipients handle surfaces previously contaminated by infected donors. Recipients touched their nasal and conjunctival mucosa after touching the surfaces. Five (50%) of 10 recipients developed infection after exposure to virus-contaminated coffee cup handles and nine (56%) of 16 became infected after exposure to contaminated plastic tiles. Spraying of contaminated tiles with a commercially available phenol/alcohol disinfectant reduced (p = 0.003) the rate of recovery of virus from the tiles from 42% (20/47) to 8% (2/26). Similarly, the rate of detection of virus on fingers touching the tiles was reduced (p = 0.001) from 61% (28/46) with unsprayed tiles to 21% (11/53) with sprayed tiles. Fifty-six per cent (9/16) of the recipients exposed on three consecutive days to untreated tiles became infected while 35% (7/20) touching only sprayed tiles became infected with rhinovirus (p = 0.3). These studies indicate that experimental rhinovirus colds can be spread by way of contaminated environmental surfaces and suggest that disinfectant treatment of such surfaces may reduce risk of viral transmission by this route.
Salinas-Carmona, M C
Nocardia brasiliensis is a Gram-positive bacterium that lives as a saprophyte in soil. In this article the physical properties, chemical composition and taxonomic position of this species is reviewed. Human infections and an experimental model of actinomycetoma in BALB/c mice as well as the host-immune response is described.
Papasian, Christopher J; Qureshi, Nilofer; Morrison, David C
The potentially protective role of the host adrenal-glucocorticoid response to enterococcal infection was evaluated in an experimental model in which mice were infected intraperitoneally with two distinct Enterococcus faecalis strains (K9 and CP-1). We demonstrated that corticosterone levels in serum and peritoneal-lavage fluid were elevated within 1 hour of infection with either E. faecalis strain. We also demonstrated that adrenalectomized mice generated a more robust localized peritoneal tumor necrosis factor alpha (TNF-alpha) response to both E. faecalis strains than did sham-adrenalectomized mice but that neither E. faecalis strain induced a systemic TNF-alpha response. Further, peritoneal TNF-alpha production in adrenalectomized mice infected with either E. faecalis K9 or CP-1 was suppressed by prior treatment with an exogenous glucocorticoid (dexamethasone). The potential clinical significance of these results was suggested by our findings that adrenalectomy markedly increased susceptibility (a>100-fold decrease in the 50% lethal dose) to lethal infections with E. faecalis CP-1 and that prior dexamethasone treatment partially compensated for adrenalectomy. In marked contrast to these findings, however, adrenalectomy did not substantially increase susceptibility to lethal E. faecalis K9 infection. Further, preinfection with E. faecalis CP-1 1 hour before infection with E. faecalis K9 did not protect mice from lethal E. faecalis K9 infections. Collectively, these studies indicate that the host can generate a glucocorticoid response to E. faecalis infection that suppresses TNF-alpha production. Further, this glucocorticoid response can protect the host from potentially lethal E. faecalis infections, but different strains show heterogeneity with respect to the extent of protection afforded by the adrenal-glucocorticoid response.
Ismail, H I H; Ashour, D S; Abou Rayia, D M; Ali, A L
Ocular lesions have been reported in patients with schistosomiasis; however, the problem with studying schistosomal infection of the human eye is that biopsies are almost impossible to take, and histopathological examination of suspicious lesions can only be undertaken post-mortem or after enucleation. This work aimed to study the possible effects and pathogenesis of schistosomiasis on the eye. This study involved 55 hamsters; five hamsters remained non-infected and the remaining 50 hamsters were infected with Schistosoma mansoni cercariae. Infected hamsters were sacrificed on weeks 8, 12, 16 and 20 post-infection (pi). Eye sections were prepared and stained for histopathological and immunohistochemical studies. Histopathological changes detected in hamsters infected after 16 and 20 weeks included looseness and oedema of the innermost retinal layers together with hyperplastic polypoid growth. Neither eggs nor granulomata were detected in eye sections throughout the experimental period. Deposition of S. mansoni antigen was revealed in 35% of infected hamsters. Later, on weeks 16 and 20 pi, moderate subepithelial conjuctival deposits and marked subchoroidal and scleral deposition were detected. In conclusion, the deposition of schistosomal antigen and immune complexes may play a pivotal role in the ocular changes that occur in schistosomiasis, even in the absence of detectable Schistosoma eggs. Schistosomiasis should be suspected in cases with unexplained ophthalmological findings, especially in endemic areas.
Gallardo, C; Soler, A; Nieto, R; Cano, C; Pelayo, V; Sánchez, M A; Pridotkas, G; Fernandez-Pinero, J; Briones, V; Arias, M
An experimental infection was conducted to evaluate horizontal transmission, clinical, virological and humoral response induced in domestic pigs infected with African swine fever (ASF) genotype II virus circulating in 2014 into the European Union (EU). Ten naive pigs were placed in contact with eight pigs experimentally inoculated with the Lithuanian LT14/1490 ASF virus (ASFV) responsible for the first ASF case detected in wild boar in Lithuania in January 2014. Clinical examination and rectal temperature were recorded each day. Blood sampling from every animal was carried out twice weekly. Blood samples were examined for presence of ASF virus-specific antibodies and for determining the ASFV viral load. From the obtained results, it was concluded that the Lithuanian ASFV induced an acute disease which resulted in 94, 5% mortality. The disease was easily detected by real-time PCR prior to the onset of clinical signs and 33% of the animals seroconverted. All findings were in accordance with observations previously made in domestic pigs and wild boar when infected with ASF genotype II viruses characterized by a high virulence. One in-contact pig remained asymptomatic and survived the infection. The role of such animals in virus transmission would need further investigation.
Meunier, Françoise; Lukan, Colette
The First European Conference on Infections in Leukaemia - ECIL1 - was organised under the auspices of the Infectious Diseases Group of the European Organisation for Research and Treatment of Cancer (EORTC), the Infectious Diseases Working Party of the European Group for Blood and Bone Marrow Transplantation (EBMT), the Supportive Care Group of the European LeukaemiaNet (ELN) and the International Immunocompromised Host Society (ICHS). The objective of the meeting was to develop evidence-based guidelines for the management of bacterial and fungal infections in high-risk immunocompromised adult leukaemia patients and hematopoietic stem cell transplantation recipients. The conference was held on September 30th and October 1st, 2005 in Juan-les-Pins, France and brought together a panel of 59 expert haematologists, oncologists, microbiologists, infectious disease specialists and clinical trialists from across Europe, Israel and Australia. The ECIL1 Guidelines were formulated after lengthy discussion, debate and panel consensus on the findings from a relevant comprehensive literature search, results of a European current practice questionnaire and other international guidelines, specific to each of the six clinical areas examined. The final recommendations, published in the Supplements of this journal as a series of six manuscripts in 2007, were well received by the medical community. The ECIL1 organisers anticipated the need for regular review of these guidelines and the Second ECIL Conference was held in September 2007. Publication of the updated and expanded ECIL2 Guidelines is forthcoming. This paper provides a concise summary of the methodology and main recommendations of the ECIL1 Guidelines.
Martín, Verónica; Mavian, Carla; López Bueno, Alberto; de Molina, Antonio; Díaz, Eduardo; Andrés, Germán; Alcami, Antonio
Amphibian-like ranaviruses include pathogens of fish, amphibians, and reptiles that have recently evolved from a fish-infecting ancestor. The molecular determinants of host range and virulence in this group are largely unknown, and currently fish infection models are lacking. We show that European sheatfish virus (ESV) can productively infect zebrafish, causing a lethal pathology, and describe a method for the generation of recombinant ESV, establishing a useful model for the study of fish ranavirus infections. PMID:26246565
Atkinson, Carter T.; Dusek, Robert J.; Woods, K.L.; Iko, W.M.
The introduction of avian malaria (Plasmodium relictum) and mosquitoes (Culex quinquefasciatus) to the Hawaiian Islands (USA) is believed to have played a major role in the decline and extinction of native Hawaiian honeycreepers (Drepanidinae). This introduced disease is thought to be one of the primary factors limiting recovery of honeycreepers at elevations below 1,200 m where native forest habitats are still relatively intact. One of the few remaining species of honeycreepers with a wide elevational distribution is the Hawaii Amakihi (Hernignathus virens). We measured morbidity and mortality in experimentally-infected Hawaii Amakihi that were captured in a high elevation, xeric habitat that is above the current range of the mosquito vector. Mortality among amakihi exposed to a single infective mosquito bite was 65% (13/20). All infected birds had significant declines in food consumption and a corresponding loss in body weight over the 60 day course of the experiment. Gross and microscopic lesions in birds that succumbed to malaria included enlargement and discoloration of the spleen and liver and parasitemias as high as 50% of circulating erythrocytes. Mortality in experimentally-infected amakihi was similar to that observed in Apapane (Himnatione sanguinea) and lower than that observed in Iiwi (Vestiaria coccinea) infected under similar conditions with the same parasite isolate. We conclude that the current elevational and geographic distribution of Hawaiian honeycreepers is determined by relative susceptibility to avian malaria.
Decaro, Nicola; Mari, Viviana; Lucente, Maria Stella; Sciarretta, Rossana; Moreno, Ana; Armenise, Carlo; Losurdo, Michele; Camero, Michele; Lorusso, Eleonora; Cordioli, Paolo; Buonavoglia, Canio
To date, limited information is available on the ability of 'Hobi'-like pestiviruses (putative bovine viral diarrhoea 3) to infect and cause disease in animal species traditionally affected by pestiviruses. In order to obtain new insights into host range and pathogenic potential of this atypical pestivirus, BVDV-seronegative calves (n=5), lambs (n=5) and piglets (n=5) were experimentally infected with the European 'Hobi'-like strain Italy-1/10-1, whereas two animals per species served as uninfected controls. Appearance of clinical signs, leukopenia, viremia, viral shedding and seroconversion were monitored for 28 days post-infection. Calves and lambs were successfully infected, displaying respiratory signs (nasal discharge), moderate hyperthermia and leukopenia, viremia and viral shedding through the nasal and faecal routes. Antibody responses were observed in both animal species by ELISA and virus neutralisation assays. In contrast, inoculated piglets did not display any clinical signs nor leukopenia and viral RNA was not detected in any biological samples. Nevertheless, the presence of detectable antibodies by virus neutralisation accounted for a successful, albeit limited infection of these animals.
Baneth, G; Shkap, V; Samish, M; Pipano, E; Savitsky, I
Canine hepatozoonosis is a disease caused by the tick-borne protozoan Hepatozoon canis. Five puppies were inoculated by ingestion of Rhipicephalus sanguineus ticks experimentally infected with H. canis, and all became infected with H. canis: gametocytes were detected in blood smears from four dogs and schizonts were observed in the spleen and bone marrow of the fifth. Antibodies reactive with H. canis gametocytes were detected by the indirect fluorescent antibody test (IFA), with IgM detected initially in all dogs 16 to 39 days post infection (PI) and IgG 22 to 43 days PI. The presence of gametocytes was first observed within peripheral blood neutrophils in Giemsa-stained blood smears between days 28 and 43 PI. Gametocyte-reactive antibodies were detected before the appearance of blood gametocytes in three of the four parasitemic dogs and also in a dog with no observed parasitemia. The detection of serum antibodies prior to the detection of blood gametocytes, or without apparent parasitemia, suggests that antibodies reactive with gametocytes may be formed against earlier forms of the parasite developing in the parenchymal tissues. Sera of dogs experimentally infected with Babesia canis, Babesia gibsoni and Ehrlichia canis exhibited no reactivity when tested with H. canis antigen. Additionally, sera positive for H. canis were not reactive with antigens of Toxoplasma gondii, Neospora caninum, Leishmania donovani and E. canis. In conclusion, incoculation of dogs with ticks infected with H. canis results in production of antibodies reactive with peripheral blood gametocytes. Detection of IgG titres would be beneficial for the diagnosis of progressive infections with undetectable parasitemia, for seroprevalence studies, and as an adjunct to IgM titres in early infections.
Tokiwa, Toshihiro; Tsugo, Kosuke; Nakamura, Shohei; Taira, Kensuke; Une, Yumi
Roundworms of the genus Baylisascaris are natural parasites primarily of wild carnivores, and they can occasionally cause infection in humans and animals. Infection results in visceral larva migrans and/or neural larva migrans, which can be severe or fatal in some animals. Recently, Baylisascaris nematodes isolated from kinkajous (Potos flavus) and previously referred to as Baylisascaris procyonis were renamed as Baylisascaris potosis; however, data regarding the pathogenicity of B. potosis towards animals and humans are lacking. In the present study, we experimentally infected squirrel monkeys (Saimiri sciureus) with B. potosis to determine the suitability of the monkey as a primate model. We used embryonated eggs of B. potosis at two different doses (10,000 eggs and 100,000 eggs) and examined the animals at 30 days post-infection. Histopathological examination showed the presence of B. potosis larvae and infiltration of inflammatory cells around a central B. potosis larvae in the brain, intestines, and liver. Nevertheless, the monkeys showed no clinical signs associated with infection. Parasitological examination revealed the presence of B. potosis larvae in the intestines, liver, lung, muscles, brain, kidney, and diaphragm. Our findings extend the range of species that are susceptible to B. potosis and provide evidence for the zoonotic potential of larva migrans in high dose infections.
Santos, Carla Domingues; Toldo, Míriam Paula Alonso; Santello, Fabrícia Helena; Filipin, Marina Del Vecchio; Brazão, Vânia; do Prado Júnior, José Clóvis
Dehydroepiandrosterone (DHEA) enhances immune responses against a wide range of viral, bacterial, and parasitic pathogens. In a previous study, we reported that administration of DHEA significantly decreased the numbers of blood parasites in Trypanosoma cruzi experimental infection. The present study was undertaken to determine the effectiveness of DHEA in reducing the severity of acute phase T. cruzi infection of male and female Wistar rats. Animals were treated subcutaneously with 40 mg/kg body weight/day of DHEA. The concentration of nitric oxide (NO) was determined in spleen peritoneal cavity. Interleukin-2 (IL-2) and interferon-gamma (IFN-gamma) were determined in the sera of uninfected and infected animals. DHEA treatment augments NO production for both sexes after in vitro LPS treatment for uninfected animals. Infection triggered enhanced NO levels although not significant. IL-2 and IFN-gamma were detectable in higher concentrations in treated and infected rats of both genders when compared to untreated controls. These data suggest that DHEA may have a potent immunoregulatory function that can affect the course of T. cruzi infection.
Sin, Yung Wa; Annavi, Geetha; Dugdale, Hannah L; Newman, Chris; Burke, Terry; MacDonald, David W
Pathogen-mediated selection is thought to maintain the extreme diversity in the major histocompatibility complex (MHC) genes, operating through the heterozygote advantage, rare-allele advantage and fluctuating selection mechanisms. Heterozygote advantage (i.e. recognizing and binding a wider range of antigens than homozygotes) is expected to be more detectable when multiple pathogens are considered simultaneously. Here, we test whether MHC diversity in a wild population of European badgers (Meles meles) is driven by pathogen-mediated selection. We examined individual prevalence (infected or not), infection intensity and co-infection of 13 pathogens from a range of taxa and examined their relationships with MHC class I and class II variability. This population has a variable, but relatively low, number of MHC alleles and is infected by a variety of naturally occurring pathogens, making it very suitable for the investigation of MHC-pathogen relationships. We found associations between pathogen infections and specific MHC haplotypes and alleles. Co-infection status was not correlated with MHC heterozygosity, but there was evidence of heterozygote advantage against individual pathogen infections. This suggests that rare-allele advantages and/or fluctuating selection, and heterozygote advantage are probably the selective forces shaping MHC diversity in this species. We show stronger evidence for MHC associations with infection intensity than for prevalence and conclude that examining both pathogen prevalence and infection intensity is important. Moreover, examination of a large number and diversity of pathogens, and both MHC class I and II genes (which have different functions), provide an improved understanding of the mechanisms driving MHC diversity.
Pietschmann, Jana; Guinat, Claire; Beer, Martin; Pronin, Valery; Tauscher, Kerstin; Petrov, Anja; Keil, Günther; Blome, Sandra
In 2007, African swine fever virus (ASFV) was introduced into the Transcaucasian countries and Russia. Since then, it has spread alarmingly and reached the European Union. ASFV strains are highly virulent and lead to almost 100% mortality under experimental conditions. However, the possibility of dose-dependent disease courses has been discussed. For this reason, a study was undertaken to assess the risk of chronic disease and the establishment of carriers upon low-dose oronasal infection of domestic pigs and European wild boar. It was demonstrated that very low doses of ASFV are sufficient to infect especially weak or runted animals by the oronasal route. Some of these animals did not show clinical signs indicative of ASF, and they developed almost no fever. However, no changes were observed in individual animal regarding the onset, course and outcome of infection as assessed by diagnostic tests. After amplification of ASFV by these animals, pen- and stablemates became infected and developed acute lethal disease with similar characteristics in all animals. Thus, we found no indication of prolonged or chronic individual courses upon low-dose infection in either species. The scattered onset of clinical signs and pathogen detection within and among groups confirms moderate contagiosity that is strongly linked with blood contact. In conclusion, the prolonged course at the "herd level" together with the exceptionally low dose that proved to be sufficient to infect a runted wild boar could be important for disease dynamics in wild-boar populations and in backyard settings.
Kaufmann, Joshka; Lenz, Tobias L; Milinski, Manfred; Eizaguirre, Christophe
Forces shaping an individual's phenotype are complex and include transgenerational effects. Despite low investment into reproduction, a father's environment and phenotype can shape its offspring's phenotype. Whether and when such paternal effects are adaptive, however, remains elusive. Using three-spined sticklebacks in controlled infection experiments, we show that sperm deficiencies in exposed males compared to their unexposed brothers functionally translated into reduced reproductive success in sperm competition trials. In non-competitive fertilisations, offspring of exposed males suffered significant costs of reduced hatching success and survival but they reached a higher body condition than their counterparts from unexposed fathers after experimental infection. Interestingly, those benefits of paternal infection did not result from increased resistance but from increased tolerance to the parasite. Altogether, these results demonstrate that parasite resistance and tolerance are shaped by processes involving both genetic and non-genetic inheritance and suggest a context-dependent adaptive value of paternal effects. PMID:25168056
van Benthem, B H B; Spaargaren, J; van den Hoek, J A R; Merks, J; Coutinho, R; Prins, M
Objectives: To investigate the prevalence and risk factors of HSV-1 and HSV-2 antibodies in HIV infected women and the association between recurrent genital ulcerations and HIV disease progression in HSV-2 positive women. Methods: The presence of HSV antibodies was tested in 276 of the 487 women participating in a European cohort study of HIV infected women. Prevalence rate ratios described the association between HSV infection and its risk factors, using log binomial regression. Generalised estimating equations (GEE) analysis was performed to determine the impact of markers of HIV disease progression on recurrent genital ulcerations. Results: The prevalence of HSV-1 and HSV-2 antibodies was 76% (95% confidence interval (95% CI): 71–81) and 42% (95% CI: 36–50); 30% (95% CI: 24–35) of the women had antibodies against both HSV-1 and HSV-2. The prevalence of HSV-1 was 86% (95% CI: 80–92) in southern Europe compared with 69% (95% CI: 57–79) and 67% (95% CI: 55–77) in central and northern Europe (p=0.002). This geographical variation remained after adjustment for other risk factors. An increasing number of years of sexual activity (p=0.0002) and a history of prostitution (p=0.0001) were independently associated with HSV-2 prevalence. In HSV-2 positive women, symptomatic cases of HSV infection were minimal, but increased with decreasing CD4 count. Conclusion: In HIV infected women, the prevalence of HSV antibodies is high and symptomatic cases of HSV infection are minimal, but increase with decreasing CD4 count. HSV-2 but not HSV-1 was related to sexual behaviour (that is, a history of prostitution and the number of sexually active years) in this group of HIV infected women. Key Words: herpes simplex viruses; genital ulcerations; HIV infection; women; Europe PMID:11287691
Dukaczewska, Agata; Tedesco, Roberto; Liesenfeld, Oliver
Ocular toxoplasmosis is a vision-threatening disease and the major cause of posterior uveitis worldwide. In spite of the continuing global burden of ocular toxoplasmosis, many critical aspects of disease including the therapeutic approach to ocular toxoplasmosis are still under debate. To assist in addressing many aspects of the disease, numerous experimental models of ocular toxoplasmosis have been established. In this article, we present an overview on in vitro, ex vivo, and in vivo models of ocular toxoplasmosis available to date. Experimental studies on ocular toxoplasmosis have recently focused on mice. However, the majority of murine models established so far are based on intraperitoneal and intraocular infection with Toxoplasma gondii. We therefore also present results obtained in an in vivo model using peroral infection of C57BL/6 and NMRI mice that reflects the natural route of infection and mimics the disease course in humans. While advances have been made in ex vivo model systems or larger animals to investigate specific aspects of ocular toxoplasmosis, laboratory mice continue to be the experimental model of choice for the investigation of ocular toxoplasmosis. PMID:26716018
Dukaczewska, Agata; Tedesco, Roberto; Liesenfeld, Oliver
Ocular toxoplasmosis is a vision-threatening disease and the major cause of posterior uveitis worldwide. In spite of the continuing global burden of ocular toxoplasmosis, many critical aspects of disease including the therapeutic approach to ocular toxoplasmosis are still under debate. To assist in addressing many aspects of the disease, numerous experimental models of ocular toxoplasmosis have been established. In this article, we present an overview on in vitro, ex vivo, and in vivo models of ocular toxoplasmosis available to date. Experimental studies on ocular toxoplasmosis have recently focused on mice. However, the majority of murine models established so far are based on intraperitoneal and intraocular infection with Toxoplasma gondii. We therefore also present results obtained in an in vivo model using peroral infection of C57BL/6 and NMRI mice that reflects the natural route of infection and mimics the disease course in humans. While advances have been made in ex vivo model systems or larger animals to investigate specific aspects of ocular toxoplasmosis, laboratory mice continue to be the experimental model of choice for the investigation of ocular toxoplasmosis.
Rivera-Benitez, José Francisco; Cuevas-Romero, Sandra; Pérez-Torres, Armando; Reyes-Leyva, Julio; Hernández, Jesús; Ramírez-Mendoza, Humberto
The aim of this study was to analyze the pathogenicity and distribution of Porcine rubulavirus (PorPV) in the respiratory tract of experimentally infected pigs. Nine 6-week-old pigs were infected with PorPV and examined clinically. Blood, nasal swab, and tissue samples were collected on different days post-infection (DPI). The humoral immune responses and viral loads were evaluated. The infected pigs exhibited an increase in the respiratory clinical signs. In addition, the excretion of PorPV was extended to 23 DPI in the nasal fluid. The distribution of PorPV in the respiratory tract tissues was extended until the end of the experiment; soft palate tonsil and lymph nodes exhibited high viral loads. The major microscopic lesions observed in the lungs corresponded to interstitial pneumonia and hyperplasia of the associated lymphoid tissue. In conclusion, PorPV infection causes a pneumonic disease characterized by a prolonged virus excretion and high viral load in the lymphoid tissues.
Castillo-Neyra, Ricardo; Borrini Mayorí, Katty; Salazar Sánchez, Renzo; Ancca Suarez, Jenny; Xie, Sherrie; Náquira Velarde, Cesar; Levy, Michael Z
Guinea pigs are important reservoirs of Trypanosoma cruzi, the causative parasite of Chagas disease, and in the Southern Cone of South America, transmission is mediated mainly by the vector Triatoma infestans. Interestingly, colonies of Triatoma infestans captured from guinea pig corrals sporadically have infection prevalence rates above 80%. Such high values are not consistent with the relatively short 7-8 week parasitemic period that has been reported for guinea pigs in the literature. We experimentally measured the infectious periods of a group of T. cruzi-infected guinea pigs by performing xenodiagnosis and direct microscopy each week for one year. Another group of infected guinea pigs received only direct microscopy to control for the effect that inoculation by triatomine saliva may have on parasitemia in the host. We observed infectious periods longer than those previously reported in a number of guinea pigs from both the xenodiagnosis and control groups. While some guinea pigs were infectious for a short time, other "super-shedders" were parasitemic up to 22 weeks after infection, and/or positive by xenodiagnosis for a year after infection. This heterogeneity in infectiousness has strong implications for T. cruzi transmission dynamics and control, as super-shedder guinea pigs may play a disproportionate role in pathogen spread.
Berra, H H; Piaggio, E; Revelli, S S; Luquita, A
Microcirculatory alterations would explain focal lesions found in Chagas' cardiomyopathy. Trypanosoma cruzi (T. cruzi) infection induces host blood properties modifications and defensive responses capable of producing blood hyperviscosity, an ischemic risk factor able to affect microvascular blood flow. We studied whole blood viscosity (eta(b)) and plasmatic and cellular factors influencing it in rats, 7 and 14 days after experimental infection with T. cruzi. Increased plasma viscosity (eta(p)) was found in infected versus control rats and it was correlated with high blood parasite levels at 7 days and enhanced gamma-globulin fraction concentration at 14 days. The hematocrit, mean corpuscular volume (MCV) and eta(b) were higher in 14 days infected rats vs. 7 days and control animals. Also, electron microscopy observation showed morphological changes in red blood cells (RBC) at 7 and 14 days post-infection, with increased proportion of echinocyte and stomatocyte shapes transformation. In our rat model of Chagas' disease, BPL, increased plasmatic protein concentration, enhanced MCV and RBC shapes transformation would determine blood hyperviscosity, cause of microvascular blood flow abnormalities.
Polverino, Eva; Bothamley, Graham H.; Goletti, Delia; Heyckendorf, Jan; Aliberti, Stefano
The breadth and quality of scientific presentations on clinical and translational research into respiratory infections at the 2015 European Respiratory Society (ERS) International Congress in Amsterdam, the Netherlands, establishes this area as one of the leadings fields in pulmonology. The host–pathogen relationship in chronic obstructive pulmonary disease, and the impact of comorbidities and chronic treatment on clinical outcomes in patients with pneumonia were studied. Various communications were dedicated to bronchiectasis and, in particular, to different prognostic and clinical aspects of this disease, including chronic infection with Pseudomonas and inhaled antibiotic therapy. Recent data from the World Health Organization showed that Europe has the highest number of multidrug-resistant tuberculosis cases and the poorest countries have the least access to suitable treatments. Latent tuberculosis and different screening programmes were also discussed with particular attention to risk factors such as HIV infection and diabetes. Several biomarkers were proposed to distinguish between active tuberculosis and latent infection. Major treatment trials were discussed (REMOX, RIFQUIN and STREAM). The possibility of once-weekly treatment in the continuation phase (RIAQUIN) was especially exciting. The continuing rise of Mycobacterium abscessus as a significant pathogen was noted. This article reviews some of the best contributions from the Respiratory Infections Assembly to the 2015 ERS International Congress. PMID:27730203
Polverino, Eva; Bothamley, Graham H; Goletti, Delia; Heyckendorf, Jan; Sotgiu, Giovanni; Aliberti, Stefano
The breadth and quality of scientific presentations on clinical and translational research into respiratory infections at the 2015 European Respiratory Society (ERS) International Congress in Amsterdam, the Netherlands, establishes this area as one of the leadings fields in pulmonology. The host-pathogen relationship in chronic obstructive pulmonary disease, and the impact of comorbidities and chronic treatment on clinical outcomes in patients with pneumonia were studied. Various communications were dedicated to bronchiectasis and, in particular, to different prognostic and clinical aspects of this disease, including chronic infection with Pseudomonas and inhaled antibiotic therapy. Recent data from the World Health Organization showed that Europe has the highest number of multidrug-resistant tuberculosis cases and the poorest countries have the least access to suitable treatments. Latent tuberculosis and different screening programmes were also discussed with particular attention to risk factors such as HIV infection and diabetes. Several biomarkers were proposed to distinguish between active tuberculosis and latent infection. Major treatment trials were discussed (REMOX, RIFQUIN and STREAM). The possibility of once-weekly treatment in the continuation phase (RIAQUIN) was especially exciting. The continuing rise of Mycobacterium abscessus as a significant pathogen was noted. This article reviews some of the best contributions from the Respiratory Infections Assembly to the 2015 ERS International Congress.
Campigotto, Gabriela; Volpato, Andréia; Galli, Gabriela M; Glombowsky, Patrícia; Baldissera, Matheus D; Miletti, Luiz Claudio; Jaguezeski, Antonise M; Stefani, Lenita M; Da Silva, Aleksandro S
Many reproductive problems has been described in male and female animals infected by Trypanosoma evansi. Thus, the aim of this study was to evaluate the occurrence of vertical (Experiment I) and venereal (Experiment II) transmission of T. evansi in rats experimentally infected. In the experiment I, eight female Wistar rats were used: three animals as negative controls, and five rats were infected by T. evansi on day ten of gestation. Out of these eight females, half puppies were used for molecular analysis (polymerase chain reaction - PCR) for T. evansi. Two infected females showed delivery problems, such as stillbirth, and fetal death that also led to female death. Three female rats infected had normal delivery of stunted offspring at term that died 2 days after birth. Rats from the control group had normal delivery of healthy offspring. T. evansi PCR was positive for 80% (12/15) of pups in the infected group. For the experiment II, five male rats were infected by T. evansi, and monitored by blood smears to check the parasitemia level. When the male rats showed parasitemia between 2 and 5 parasites per field, they were individually housed with one female adult rat. After approximately 21 days, the females delivered their offspring. Blood sample was collected from the females for blood smears and T. evansi PCR tests, which revealed negative results. Therefore, we were able to prove the occurrence of transplacental transmission of T. evansi and its negative effect on female rats and their offspring.
Bertó-Moran, Alejandro; Pacios, Isabel; Serrano, Emmanuel; Moreno, Sacramento; Rouco, Carlos
The interaction among several parasites in European rabbits (Oryctolagus cuniculus) is crucial to host fitness and to the epidemiology of myxomatosis and rabbit hemorrhagic disease. These diseases have caused significant reductions in rabbit populations on the Iberian Peninsula. Most studies have focused on the epidemiology and pathogenesis of these viruses individually, and little is known about interactions between these viruses and other parasites. Taking advantage of an experimental restocking program in Spain, the effects of coccidian and nematode infections on the probability of having detectable antibody to myxoma and rabbit hemorrhagic disease viruses were tested in European wild rabbits. For 14 mo, we monitored rabbit abundance and parasite loads (coccidia and nematodes) in three reintroduced rabbit populations. While coccidian and nematode loads explained seasonal antibody prevalences to myxoma virus, the pattern was less clear for rabbit hemorrhagic disease. Contrary to expectations, prevalence of antibody to myxoma virus was inversely proportional to coccidian load, while nematode load seemed to play a minor role. These results have implications for viral disease epidemiology and for disease management intended to increase rabbit populations in areas where they are important for ecosystem conservation.
Gervasi, G; Capanna, A; Mita, V; Zaratti, L; Franco, E
Rotavirus (RV) is worldwide considered as the most important viral agent of acute gastroenteritis in children less than 5 y. Since 2006, the availability of anti-RV vaccines has deeply modified the incidence and economic burden of RV infection. In Europe, some countries have introduced an anti-RV vaccination program in the last 10 y. Although community acquired RV (CARV) disease is the most studied condition of RV infection, recently some authors have highlighted the importance of nosocomial RV (nRV) disease as an emerging public health issue. The aim of this review is to summarize the epidemiology of both CARV and nRV, in order to discuss the difficulty of a clear evaluation of the burden of the disease in absence of comparable data. In particular, we focused our attention to European studies regarding nRV in terms of divergences related to definition, report of incidence rate and methodological issues.
Mühldorfer, Kristin; Speck, Stephanie; Kurth, Andreas; Lesnik, René; Freuling, Conrad; Müller, Thomas; Kramer-Schadt, Stephanie; Wibbelt, Gudrun
Background Bats receive increasing attention in infectious disease studies, because of their well recognized status as reservoir species for various infectious agents. This is even more important, as bats with their capability of long distance dispersal and complex social structures are unique in the way microbes could be spread by these mammalian species. Nevertheless, infection studies in bats are predominantly limited to the identification of specific pathogens presenting a potential health threat to humans. But the impact of infectious agents on the individual host and their importance on bat mortality is largely unknown and has been neglected in most studies published to date. Methodology/Principal Findings Between 2002 and 2009, 486 deceased bats of 19 European species (family Vespertilionidae) were collected in different geographic regions in Germany. Most animals represented individual cases that have been incidentally found close to roosting sites or near human habitation in urban and urban-like environments. The bat carcasses were subjected to a post-mortem examination and investigated histo-pathologically, bacteriologically and virologically. Trauma and disease represented the most important causes of death in these bats. Comparative analysis of pathological findings and microbiological results show that microbial agents indeed have an impact on bats succumbing to infectious diseases, with fatal bacterial, viral and parasitic infections found in at least 12% of the bats investigated. Conclusions/Significance Our data demonstrate the importance of diseases and infectious agents as cause of death in European bat species. The clear seasonal and individual variations in disease prevalence and infection rates indicate that maternity colonies are more susceptible to infectious agents, underlining the possible important role of host physiology, immunity and roosting behavior as risk factors for infection of bats. PMID:22216354
Sandgren, Andreas; Vonk Noordegraaf-Schouten, Jannigje M; Oordt-Speets, Anouk M; van Kessel, Gerarda B; de Vlas, Sake J; van der Werf, Marieke J
Individuals with latent tuberculosis infection (LTBI) are the reservoir of Mycobacterium tuberculosis in a population and as long as this reservoir exists, elimination of tuberculosis (TB) will not be feasible. In 2013, the European Centre for Disease Prevention and Control (ECDC) started an assessment of benefits and risks of introducing programmatic LTBI control, with the aim of providing guidance on how to incorporate LTBI control into national TB strategies in European Union/European Economic Area (EU/EEA) Member States and candidate countries. In a first step, experts from the Member States, candidate countries, and international and national organisations were consulted on the components of programmatic LTBI control that should be considered and evaluated in literature reviews, mathematical models and cost-effectiveness studies. This was done through a questionnaire and two interactive discussion rounds. The main components identified were identification and targeting of risk groups, determinants of LTBI and progression to active TB, optimal diagnostic tests for LTBI, effective preventive treatment regimens, and to explore the potential for combining LTBI control with other health programmes. Political commitment, a solid healthcare infrastructure, and favourable economic situation in specific countries were identified as essential to facilitate the implementation of programmatic LTBI control. PMID:27589214
Sandgren, Andreas; Vonk Noordegraaf-Schouten, Jannigje M; Oordt-Speets, Anouk M; van Kessel, Gerarda B; de Vlas, Sake J; van der Werf, Marieke J
Individuals with latent tuberculosis infection (LTBI) are the reservoir of Mycobacterium tuberculosis in a population and as long as this reservoir exists, elimination of tuberculosis (TB) will not be feasible. In 2013, the European Centre for Disease Prevention and Control (ECDC) started an assessment of benefits and risks of introducing programmatic LTBI control, with the aim of providing guidance on how to incorporate LTBI control into national TB strategies in European Union/European Economic Area (EU/EEA) Member States and candidate countries. In a first step, experts from the Member States, candidate countries, and international and national organisations were consulted on the components of programmatic LTBI control that should be considered and evaluated in literature reviews, mathematical models and cost-effectiveness studies. This was done through a questionnaire and two interactive discussion rounds. The main components identified were identification and targeting of risk groups, determinants of LTBI and progression to active TB, optimal diagnostic tests for LTBI, effective preventive treatment regimens, and to explore the potential for combining LTBI control with other health programmes. Political commitment, a solid healthcare infrastructure, and favourable economic situation in specific countries were identified as essential to facilitate the implementation of programmatic LTBI control.
Kraase, Martin; Sloan, Richard; Klein, Dieter; Logan, Nicola; McMonagle, Linda; Biek, Roman; Willett, Brian J; Hosie, Margaret J
Feline immunodeficiency virus (FIV), an immunosuppressive lentivirus found in cats worldwide, is studied to illuminate mechanisms of lentiviral pathogenesis and to identify key components of protective immunity. During replication, lentiviruses accumulate errors of nucleotide mis-incorporation due to the low-fidelity of reverse transcriptase and recombination between viral variants, resulting in the emergence of a complex viral "quasispecies". In patients infected with HIV-1, env sequences may vary by up to 10% and the detection of quasispecies with greater heterogeneity is associated with higher viral loads and reduced CD4+ T cell numbers , indicating that transmission of more complex quasispecies may lead to disease progression. However, little is known about how FIV evolves as disease progresses, or why some cats develop AIDS rapidly while disease progression is slow in others. The aim of this study was to determine whether disease progression may be governed by viral evolution and to examine the diversity of viral variants emerging following infection with an infectious molecular clone. The FIV env gene encoding the envelope glycoprotein (Env) was examined at early (12 weeks) and late (322 weeks) stages of FIV infection in two groups of cats infected experimentally with the FIV-GL8 molecular clone. Viral variants were detected within quasispecies in cats in the late stages of FIV infection that contained differing amino acid compositions in several variable loops of Env, some of which were identified as determinants of receptor usage and resistance to neutralization. Therefore these results indicate that the FIV env gene evolves during the course of infection, giving rise to variants that resist neutralization and likely lead to disease progression.
Ricklin, Meret E; Garcìa-Nicolàs, Obdulio; Brechbühl, Daniel; Python, Sylvie; Zumkehr, Beatrice; Posthaus, Horst; Oevermann, Anna; Summerfield, Artur
Pigs are considered to be the main amplifying host for Japanese encephalitis virus (JEV), and their infection can correlate with human cases of disease. Despite their importance in the ecology of the virus as it relates to human cases of encephalitis, the pathogenesis of JEV in pigs remains obscure. In the present study, the localization and kinetics of virus replication were investigated in various tissues after experimental intravenous infection of pigs. The data demonstrate a rapid and broad spreading of the virus to the central nervous system (CNS) and various other organs. A particular tropism of JEV in pigs not only to the CNS but also for secondary lymphoid tissue, in particular the tonsils with the overall highest viral loads, was observed. In this organ, even 11 days post infection, the latest time point of the experiment, no apparent decrease in viral RNA loads and live virus was found despite the presence of a neutralizing antibody response. This was also well beyond the clinical and viremic phase. These results are of significance for the pathogenesis of JEV, and call for further experimental studies focusing on the cellular source and duration of virus replication in pigs.
Howe, George E.; Stehly, Guy R.
A method was developed to experimentally induce saprolegniasis in rainbow trout Oncorhynchus mykiss. The development of a reliable method to produce infected fish is essential to efforts to determine the efficacy of various antifungal treatments. Three methods for inducing saprolegniasis were evaluated in waters containing known concentrations of Saprolegnia parasitica zoospores. These methods included application of the following stressors to fish: (1) abrasion and dewatering, (2) water temperature increase, and (3) a combination of abrasion, dewatering, and temperature increase. Neither physical abrasion nor temperature increase stress alone was effective for inducing saprolegniasis. Only 25.9% of fish stressed by abrasion and dewatering alone became infected. Application of both abrasion and temperature stress, however, induced saprolegniasis in 77.8% of fish tested. Most of these fish became infected after 5 d of stress treatments. No fish became infected or died in the positive control group (not stressed but exposed to S. parasitica zoospores) or the negative control group (not stressed or challenged). This method should enable researchers to induce saprolegniasis in rainbow trout to study its pathogenesis or to test the efficacy of antifungal treatments. In conducting efficacy studies, it is important that therapeutic treatments begin promptly after the first signs of saprolegniasis are observed because the disease can progress very quickly and often results in mortality.
de Carvalho Ferreira, H C; Weesendorp, E; Quak, S; Stegeman, J A; Loeffen, W L A
Knowledge on African Swine Fever (ASF) transmission routes can be useful when designing control measures against the spread of ASF virus (ASFV). Few studies have focused on the airborne transmission route, and until now no data has been available on quantities of ASF virus (ASFV) in the air. Our aim was to validate an air sampling technique for ASF virus (ASFV) that could be used to detect and quantify virus excreted in the air after experimental infection of pigs. In an animal experiment with the Brazil'78, the Malta'78 and Netherlands'86 isolates, air samples were collected at several time points. For validation of the air sampling technique, ASFV was aerosolised in an isolator, and air samples were obtained using the MD8 air scan device, which was shown to be suitable to detect ASFV. The half-life of ASFV in the air was on average 19 min when analysed by PCR, and on average 14 min when analysed by virus titration. In rooms with infected pigs, viral DNA with titres up to 10(3.2) median tissue culture infective dose equivalents (TCID50eq.)/m(3) could be detected in air samples from day 4 post-inoculation (dpi 4) until the end of the experiments, at dpi 70. In conclusion, this study shows that pigs infected with ASFV will excrete virus in the air, particularly during acute disease. This study provides the first available parameters to model airborne transmission of ASFV.
Da Silva, Aleksandro Schafer; Costa, Márcio Machado; Wolkmer, Patrícia; Zanette, Régis Adriel; Faccio, Luciana; Gressler, Lucas Trevisan; Dorneles, Tagor Eduardo Andreolla; Santurio, Janio Morais; Lopes, Sonia Terezinha dos Anjos; Monteiro, Silvia Gonzalez
This study aimed at evaluating hemogram and erythropoietic changes in cats experimentally infected with Trypanosoma evansi. Thirteen adult female non-breeding Felix catus were separated into two groups: seven animals were infected with 10(8) trypomastigotes each, and six animals were used as negative controls. Animals were kept in air-conditioned rooms and blood smears were performed daily for 49 days. Blood samples were collected from the jugular vein at days 0, 7, 21, 35 and 49 and stored in blood-collecting tubes containing anticoagulant. Bone marrow was collected from the proximal epiphysis of the right femur at days 14 and 42 post-inoculation (PI). Total erythrocyte count, hematocrit and hemoglobin showed statistical differences among groups from the seventh day PI onwards (P<0.05). The mean corpuscular volume and the mean corpuscular hemoglobin concentration remained normal, characterizing a normocytic-normochromic anemia. Reticulocyte count increased in the infected group from the 21st day onwards, but remained near normal values suggesting a mild regenerative anemia. Moreover, the myeloid:erythroid ratio significantly reduced at day 42 PI, evidencing a bone marrow hematopoietic response. Based on these results we conclude that cats infected with T. evansi have normocytic, normochromic, regenerative anemia.
Boulouis, Henri J.; Barrat, Francine; Bermond, Delphine; Bernex, Florence; Thibault, Danièle; Heller, Rémy; Fontaine, Jean-Jacques; Piémont, Yves; Chomel, Bruno B.
The kinetics of infection and the pathogenic effects on the reproductive function of laboratory mice infected with Bartonella birtlesii recovered from an Apodemus species are described. B. birtlesii infection, as determined by bacteremia, occurred in BALB/c mice inoculated intravenously. Inoculation with a low-dose inoculum (1.5 × 103 CFU) induced bacteremia in only 75% of the mice compared to all of the mice inoculated with higher doses (≥1.5 × 104). Mice became bacteremic for at least 5 weeks (range, 5 to 8 weeks) with a peak ranging from 2 × 103 to 105 CFU/ml of blood. The bacteremia level was significantly higher in virgin females than in males but the duration of bacteremia was similar. In mice infected before pregnancy (n = 20), fetal loss was evaluated by enumerating resorption and fetal death on day 18 of gestation. The fetal death and resorption percentage of infected mice was 36.3% versus 14.5% for controls (P < 0.0001). Fetal suffering was evaluated by weighing viable fetuses. The weight of viable fetuses was significantly lower for infected mice than for uninfected mice (P < 0.0002). Transplacental transmission of Bartonella was demonstrated since 76% of the fetal resorptions tested was culture positive for B. birtlesii. The histopathological analysis of the placentas of infected mice showed vascular lesions in the maternal placenta, which could explain the reproductive disorders observed. BALB/c mice appeared to be a useful model for studying Bartonella infection. This study provides the first evidence of reproductive disorders in mice experimentally infected with a Bartonella strain originating from a wild rodent. PMID:11500400
Ferreira, Milene Silveira; de Castro, Paulo Henrique Gomes; Silva, Gilmara Abreu; Casseb, Samir Mansur Moraes; Dias Júnior, Antônio Gregório; Rodrigues, Sueli Guerreiros; Azevedo, Raimunda do Socorro da Silva; Costa e Silva, Matheus Fernandes; Zauli, Danielle Alves Gomes; Araújo, Márcio Sobreira Silva; Béla, Samantha Ribeiro; Teixeira-Carvalho, Andréa; Martins-Filho, Olindo Assis; Vasconcelos, Pedro Fernando da Costa
Although the murine models have the feasibility to reproduce some signs of dengue Virus (DENV) infection, the use of isogenic hosts with polarized immune response patterns does not reproduce the particularities of human disease. Our goal was to investigate the kinetics of peripheral blood biomarkers in immunocompetent Callithrix penicillata non-human primates subcutaneously infected with DENV-3. The viral load of infected animals was determinated by quantitative real time PCR. Measurements of DENV-3/IgM were performed, and several parameters were assessed by hemogram: red blood cells count, hemoglobin, hematocrit, white blood cells count, neutrophils, monocytes, lymphocytes, and platelets count. The coagulogram was performed by prothrombin time (PT), and activated partial thromboplastin time (APTT) assays. The renal function was monitored by urea and creatinine, and the liver function by the aspartate (AST), and alanine (ALT) aminotransferases. Also, the level of the cytokines IL-6, TNF-α, IL-2, IFN-γ, IL-4 and IL-5 was quantified during the experimental study. Data analysis was performed considering relevant differences when baseline fold changes were found outside from 0.75 to 1.5 range. Our data demonstrated that infected animals presented relevant signs of dengue disease, including peaks of viremia at 5 days-post-infection (dpi), peaks of anti-DENV-3 IgM at 15 dpi and hemaglutination inhibition assay (HIA) from 15 to at 60 dpi. Despite early monocytosis, slight neutrophilia and lymphocytosis, animals developed persistent leucopenia starting at 4 dpi. Anemia episodes were steady at 3-4 dpi. Patent thrombocytopenia was observed from 1 to 15 dpi with sporadic decrease of APTT. A substantial increase of ALT and AST was observed with higher peak at 4 dpi. Moreover, early increases of TNF-alpha and IFN-gamma besides late increase of IFN-gamma were observed. The analysis of biomarkers network pointed out two relevant strong axes during early stages of dengue fever
Ribicich, M; Krivokapich, S; Pasqualetti, M; Gonzalez Prous, C L; Gatti, G M; Falzoni, E; Aronowicz, T; Arbusti, P; Fariña, F; Rosa, A
Trichinella spiralis has been documented in wild animals in Argentina, including puma, armadillos, rats and wild boars. In 2008, molecular analysis identified Trichinella T12 from a naturally infected puma (Puma concolor) from Patagonia. The aim of the present work was to study the relationship between the infectivity and pathology of Trichinella T12 in the puma and in domestic cats, and the possible risks that may be present for transmission among these animals. Two cats (A and B) were orally-infected with 3300 and 1850 Trichinella T12 muscle larvae, respectively; one additional cat was used as a control. During the 54 days post-infection, a daily examination was performed which included monitoring body temperature, and cardiac and respiration rates; the animals were then euthanized. Hematological studies included hematocrit (%), hemoglobin (g/dl), and white cell, neutrophil, lymphocyte and eosinophil counts. Blood biochemistry included urea, creatinine, AST, ALT, CK, LDH and ALP. An ELISA assay was also performed. At necropsy, organs (liver, spleen, brain, cerebellum and kidney), nails and muscle samples were obtained for histopathology studies and artificial digestion. The muscles that were studied included the diaphragm, massetter, cutaneous, temporal, intercostals, lumbar, tongue, limbs, neck and tail. Clinical signs, such as anorexia, diarrhea, vomiting, shaggy hair, decay and muscle pain, were observed in both cats. The eosinophil counts were elevated in both cats A and B. Trichinella larvae were recovered from all of the muscles analyzed where the histopathology showed larvae in several muscles without degenerative reaction. Neither larvae nor lesions were observed in non-muscular organs. Cat A had a maximum of 246 larvae per gram (lpg) in the temporal muscle and a minimum of 80 lpg in the tongue, while cat B had a maximum of 65 lpg in muscles of the leg and a minimum of 10 lpg in tail muscles. This study represents the first record of experimental
Zaugg, J L
Anaplasma ovis was experimentally transmitted from sheep to pronghorn antelope (Antilocapra americana) and back to sheep. Anaplasma ovis was recovered in splenectomized sheep, from two of three spleen-intact pronghorns following their inoculation with blood from known A. ovis carrier sheep. These two pronghorns exhibited a 0.5% or higher A. ovis parasitemia within 48 days after exposure, and an anaplasmosis-positive serological response 91 days after exposure. Clinical signs of illness were not observed. Blood from the infected pronghorns produced disease in four splenectomized sheep.
Deardorff, Eleanor R.; Forrester, Naomi L.; Travassos da Rosa, Amelia P.; Estrada-Franco, Jose G.; Navarro-Lopez, Roberto; Tesh, Robert B.; Weaver, Scott C.
Coues rice rat (Oryzomys couesi), a species abundant throughout Central America, was evaluated experimentally for the ability to serve as an amplifying host for three arboviruses: Patois (Bunyaviridae, Orthobunyavirus), Nepuyo (Orthobunyavirus), and Venezuelan equine encephalitis virus subtype ID (Togaviridae, Alphavirus). These three viruses have similar ecologies and are known to co-circulate in nature. Animals from all three cohorts survived infection and developed viremia with no apparent signs of illness and long-lasting antibodies. Thus, O. couesi may play a role in the general maintenance of these viruses in nature. PMID:20134016
Deardorff, Eleanor R; Forrester, Naomi L; Travassos da Rosa, Amelia P; Estrada-Franco, Jose G; Navarro-Lopez, Roberto; Tesh, Robert B; Weaver, Scott C
Coues rice rat (Oryzomys couesi), a species abundant throughout Central America, was evaluated experimentally for the ability to serve as an amplifying host for three arboviruses: Patois (Bunyaviridae, Orthobunyavirus), Nepuyo (Orthobunyavirus), and Venezuelan equine encephalitis virus subtype ID (Togaviridae, Alphavirus). These three viruses have similar ecologies and are known to co-circulate in nature. Animals from all three cohorts survived infection and developed viremia with no apparent signs of illness and long-lasting antibodies. Thus, O. couesi may play a role in the general maintenance of these viruses in nature.
Samuels, David C.; Kallianpur, Asha R.; Ellis, Ronald J.; Bush, William S.; Letendre, Scott; Franklin, Donald; Grant, Igor; Hulgan, Todd
Background Mitochondrial DNA (mtDNA) haplogroups are ancestry-related patterns of single-nucleotide polymorphisms that are associated with differential mitochondrial function in model systems, neurodegenerative diseases in HIV-negative populations, and chronic complications of HIV infection, including neurocognitive impairment. We hypothesized that mtDNA haplogroups are associated with neuroinflammation in HIV-infected adults. Methods CNS HIV Antiretroviral Therapy Effects Research (CHARTER) is a US-based observational study of HIV-infected adults who underwent standardized neurocognitive assessments. Participants who consented to DNA collection underwent whole blood mtDNA sequencing, and a subset also underwent lumbar puncture. IL-6, IL-8, TNF-α (high-sensitivity), and IP-10 were measured in cerebrospinal fluid (CSF) by immunoassay. Multivariable regression of mtDNA haplogroups and log-transformed CSF biomarkers were stratified by genetic ancestry using whole-genome nuclear DNA genotyping (European [EA], African [AA], or Hispanic ancestry [HA]), and adjusted for age, sex, antiretroviral therapy (ART), detectable CSF HIV RNA, and CD4 nadir. A total of 384 participants had both CSF cytokine measures and genetic data (45% EA, 44% AA, 11% HA, 22% female, median age 43 years, 74% on ART). Results In analyses stratified by the 3 continental ancestry groups, no haplogroups were significantly associated with the 4 biomarkers. In the subgroup of participants with undetectable plasma HIV RNA on ART, European haplogroup H participants had significantly lower CSF TNF-α (P = 0.001). Conclusions Lower CSF TNF-α may indicate lower neuroinflammation in the haplogroup H participants with well-controlled HIV on ART.
Schwarz, Patrick; Bretagne, Stéphane; Gantier, Jean-Charles; Garcia-Hermoso, Dea; Lortholary, Olivier; Dromer, Françoise; Dannaoui, Eric
Mucormycosis is an emerging infection associated with a high mortality rate. Identification of the causative agents remains difficult and time-consuming by standard mycological procedures. In this study, internal transcribed spacer (ITS) sequencing was validated as a reliable technique for identification of Zygomycetes to the species level. Furthermore, species identification directly from infected tissues was evaluated in experimentally infected mice. Fifty-four Zygomycetes strains belonging to 16 species, including the most common pathogenic species of Rhizopus spp., Absidia spp., Mucor spp., and Rhizomucor spp., were used to assess intra- and interspecies variability. Ribosomal DNA including the complete ITS1-5.8S-ITS2 region was amplified with fungal universal primers, sequenced, and compared. Overall, for a given species, sequence similarities between isolates were >98%. In contrast, ITS sequences were very different between species, allowing an accurate identification of Zygomycetes to the species level in most cases. Six species (Rhizopus oryzae, Rhizopus microsporus, Rhizomucor pusillus, Mucor circinelloides, and Mucor indicus) were also used to induce disseminated mucormycosis in mice and to demonstrate that DNA extraction, amplification of fungal DNA, sequencing, and molecular identification were possible directly from frozen tissues. PMID:16455881
Gast, R K; Beard, C W
The antibody response of laying hens to experimental Salmonella enteritidis infection was evaluated in microagglutination, tube agglutination, and rapid whole-blood plate agglutination assays. Hens of three different ages were infected by either oral inoculation or horizontal contact transmission. Blood was collected at weekly intervals, and the presence of specific antibodies was assessed by reaction with antigens prepared from strains of S. enteritidis and S. pullorum. The sensitivity of detection of infected hens did not vary significantly between the assays, as all three tests effectively identified most exposed hens as seropositive. Within each test, however, variation was observed in the detection sensitivity when different antigens were used. The microagglutination titers of serum samples were determined by serial dilution. Antibody titers peaked at 1 to 2 weeks postinoculation and declined steadily, although most birds were still identified as seropositive at 10 weeks postinoculation. The mean microtest titers obtained with S. enteritidis antigens were higher than with an S. pullorum antigen, indicating greater test sensitivity. However, use of the S. pullorum antigen resulted in fewer false positives when sera from uninfected control hens were tested. The titers of contact-exposed hens peaked later and at lower values than did those of inoculated hens, but these two groups of hens had similar antibody titers after the third week postinoculation.
Woodroffe, Rosie; Donnelly, Christl A; Wei, Gao; Cox, D R; Bourne, F John; Burke, Terry; Butlin, Roger K; Cheeseman, C L; Gettinby, George; Gilks, Peter; Hedges, Simon; Jenkins, Helen E; Johnston, W Thomas; McInerney, John P; Morrison, W Ivan; Pope, Lisa C
1. In most social animals, the prevalence of directly transmitted pathogens increases in larger groups and at higher population densities. Such patterns are predicted by models of Mycobacterium bovis infection in European badgers (Meles meles). 2. We investigated the relationship between badger abundance and M. bovis prevalence, using data on 2696 adult badgers in 10 populations sampled at the start of the Randomized Badger Culling Trial. 3. M. bovis prevalence was consistently higher at low badger densities and in small social groups. M. bovis prevalence was also higher among badgers whose genetic profiles suggested that they had immigrated into their assigned social groups. 4. The association between high M. bovis prevalence and small badger group size appeared not to have been caused by previous small-scale culling in study areas, which had been suspended, on average, 5 years before the start of the current study. 5. The observed pattern of prevalence might occur through badgers in smaller groups interacting more frequently with members of neighbouring groups; detailed behavioural data are needed to test this hypothesis. Likewise, longitudinal data are needed to determine whether the size of infected groups might be suppressed by disease-related mortality. 6. Although M. bovis prevalence was lower at high population densities, the absolute number of infected badgers was higher. However, this does not necessarily mean that the risk of M. bovis transmission to cattle is highest at high badger densities, since transmission risk depends on badger behaviour as well as on badger density.
Herrera, Jenifer; Gómez-Núñez, Luis; Lara-Romero, Rocío; Diosdado, Fernando; Martínez-Lara, Atalo; Jasso, Miguel; Ramírez-Mendoza, Humberto; Pérez-Torres, Armando; Rivera-Benítez, José Francisco
The objective of this study was to evaluate the clinical disease, humoral response and viral distribution of recent Porcine rubulavirus (PorPV) isolates in experimentally infected pigs. Four, 6-piglet (5-days old) groups were employed (G1-84, G2-93, G3-147, and G4-T). Three viral strains were used for the experimental infection: the reference strain LPMV-1984 (Michoacán 1984) and two other strains isolated in 2013, one in Queretaro (Qro/93/2013) and the other in Michoacán (Mich/147/2013). Each strain was genetically characterized by amplification and sequencing of the gene encoding hemagglutinin-neuroamidase (HN). The inoculation was performed through the oronasal and ocular routes, at a dose of 1×10(6)TCID50/ml. Subsequently, the signs were evaluated daily and necropsies were performed on 3 different days post infection (dpi). We recorded all micro- and macroscopic lesions. Organs from the nervous, lymphatic, and respiratory system were analyzed by quantifying the viral RNA load and the presence of the infectious virus. The presence of the viral antigen in organs was evidenced through immunohistochemistry. Seroconversion was evaluated through the use of a hemagglutination inhibition test. In the characterization of gene HN, only three substitutions were identified in strain Mich/147/2013, two in strain LPMV/1984 (fourth passage) and one in strain Qro/93/2013, with respect to reference strain LPMV-84, these changes had not been identified as virulence factors in previously reported strains. Neurological alterations associated with the infection were found in all three experimental groups starting from 3dpi. Groups G1-84 and G3-147 presented the most exacerbated nervous signs. Group G2-93 only presented milder signs including slight motor incoordination, and an increased rectal temperature starting from day 5 post infection (PI). The main histopathological findings were the presence of a mononuclear inflammatory infiltrate (lymphocytic/monocytic) surrounding the
Somova, L M; Kondrashova, N M; Plekhova, N G; Drobot, E I; Lyapun, I N
Pathomorphological changes in the organs of animals intranasally infected with Streptococcus pneumoniae were studied under conditions of immunotropic therapy added to antibiotic therapy. The pathomorphosis in the lungs, spleen, and thymus in animals treated with likopid, tinrostim, and roncoleukin was described. A positive time course of the pathological process in experimental animals in comparison with intact animals and animals receiving no immunotropic drugs was demonstrated.
Stott, Philip; O'Callaghan, Michael; Phillips, Peter; Verbyla, Ari
The factors that control the demography of European hare Lepus europaeus populations are poorly understood, but it has been recognized that the decline of hares in Europe is associated with an increasing intensity of agricultural activity. Many mechanisms have been suggested. We propose another mechanism; a negative impact arising from ingestion of the infective larvae of ruminant livestock. We dosed juvenile hares from a worm-free colony with a conservative dose from a mixed culture of infective larvae of the nematode parasites of sheep and cattle. We examined the hares post-mortem for the establishment of those ruminant nematodes, differences in weight changes, and the shedding of eggs. We found that under the circumstances of our trial, Trichostrongylus colubriformis and to a lesser extent T. rugatus, T. vitrinus, and Teladorsagia circumcincta were able to establish as adults in the dosed animals. We found strongyle eggs in the faeces of the dosed hares, and were able to culture larvae from those eggs. However, the ecological significance of our findings, if any, remains to be elucidated. Because of their mobility, hares may transmit resistant strains of parasites between grazing properties.
Franklinos, Lydia H V; Efstratiou, Androulla; Macgregor, Shaheed K; John, Shinto K; Hopkins, Timothy; Cunningham, Andrew A; Lawson, Becki
Streptococcus pyogenes, a common pathogen of humans, was isolated from the carcass of a free-living European hedgehog (Erinaceus europaeus) found in northern England in June 2014. The animal had abscessation of the deep right cervical lymph node, mesenteric lymph nodes and liver. The S. pyogenes strain isolated from the lesions, peritoneal and pleural cavities was characterised as emm 28, which can be associated with invasive disease in humans. This is the first known report of S. pyogenes in a hedgehog and in any free-living wild animal that has been confirmed by gene sequencing. As close associations between wild hedgehogs and people in England are common, we hypothesise that this case might have resulted from anthroponotic infection.
Fort, Marcelo; Baldone, Valeria; Fuchs, Lumila; Giménez, Hugo; Rojas, María; Breccia, Javier D; Oyhenart, Jorge
Brucella suis biovar 1 is the causative agent of brucellosis in several domestic and wild animals and it is a common agent of human brucellosis. European hares (Lepus europaeus) have been shown to be infected by B. suis biovar 1 and the transmission to other animals has been suggested. In this work, experimental rabbits (Cuniculus orictolagus) were infected with B. suis biovar 1 isolated from wild hares. Infected rabbits showed high serological response in 2 weeks after discharge and typical granulomatous lesions (2mm diameter) were found in liver, spleen and kidneys after 50 days. B. suis biovar 1 was cultured from the lesion of the organs mentioned above as well as from urine, placenta and fetuses. These data suggest that hares are a potential source for horizontal transmission of B. suis biovar 1 to other mammalians.
Handeland, Kjell; Nesse, Live L; Lillehaug, Atle; Vikøren, Turid; Djønne, Berit; Bergsjø, Bjarne
The red fox (Vulpes vulpes) can be considered as a relevant indicator species for Salmonella in the local environment and Salmonella faecal carriage was investigated in 215 red foxes in Norway shot during the winters 2002/2003 and 2003/2004. Fourteen (6.5%) of the foxes carried Salmonella. Four isolates were determined as serovars Kottbus (n=2) and Hessarek (n=2) of Salmonella enterica subspecies enterica, and one as S. enterica subspecies IIIb:61:k:1,5,(7). The remaining nine isolates were S. enterica subspecies enterica serovar Typhimurium 4,12:i:1,2 and all displayed the same pulsed-field gel electrophoresis (PFGE) profile designated A2. This serovar regularly causes disease outbreaks amongst small passerine birds during winter and most of these outbreaks are associated with the PFGE profile A2. The results strongly indicated that the Salmonella Typhimurium infections in red foxes were primarily acquired through ingestion of infected small passerines. To investigate the capability of the A2 strain to establish a true intestinal infection in the fox an inoculation experiment with an A2 isolate from small passerines was carried out in farmed silver foxes (V. vulpes). The experiment also included one strain with an uncommonly occurring profile (X201) from small passerines. To highlight possible differences in capability of the two inoculation strains to pass the acid gastric juice in the fox, in vitro studies of their acid tolerance was carried out. Also their catalase activity and biofilm production were studied. All three foxes inoculated with the A2 strain developed sub-clinical intestinal infection of 2 weeks duration, whereas none of the three foxes inoculated with the X201 strain shed this bacterium. The X201 strain displayed a much lower capability, than the A2 strain, to survive at pH 3 in vitro. The low acid tolerance probably made it difficult for the X201 strain to pass the stomach and establish an intestinal infection in the experimental foxes. Reduced
Tsukiyama-Kohara, Kyoko; Kohara, Michinori
Tupaias, or tree shrews, are small mammals that are similar in appearance to squirrels. The morphological and behavioral characteristics of the group have been extensively characterized, and despite previously being classified as primates, recent studies have placed the group in its own family, the Tupaiidae. Genomic analysis has revealed that the genus Tupaia is closer to humans than it is to rodents. In addition, tupaias are susceptible to hepatitis B virus and hepatitis C virus. The only other experimental animal that has been demonstrated to be sensitive to both of these viruses is the chimpanzee, but restrictions on animal testing have meant that experiments using chimpanzees have become almost impossible. Consequently, the development of the tupaia for use as an animal infection model could become a powerful tool for hepatitis virus research and in preclinical studies on drug development. PMID:25048261
Arsnoe, D.M.; Ip, H.S.; Owen, J.C.
Migrating waterfowl are implicated in the global spread of influenza A viruses (IAVs), and mallards (Anas platyrhynchos) are considered a particularly important IAV reservoir. Prevalence of IAV infection in waterfowl peaks during autumn pre-migration staging and then declines as birds reach wintering areas. Migration is energetically costly and birds often experience declines in body condition that may suppress immune function. We assessed how body condition affects susceptibility to infection, viral shedding and antibody production in wild-caught and captive-bred juvenile mallards challenged with low pathogenic avian influenza virus (LPAIV) H5N9. Wild mallards (n = 30) were separated into three experimental groups; each manipulated through food availability to a different condition level (-20%, -10%, and normal ??5% original body condition), and captive-bred mallards (n = 10) were maintained at normal condition. We found that wild mallards in normal condition were more susceptible to LPAIV infection, shed higher peak viral loads and shed viral RNA more frequently compared to birds in poor condition. Antibody production did not differ according to condition. We found that wild mallards did not differ from captive-bred mallards in viral intensity and duration of infection, but they did exhibit lower antibody titers and greater variation in viral load. Our findings suggest that reduced body condition negatively influences waterfowl host competence to LPAIV infection. This observation is contradictory to the recently proposed condition-dependent hypothesis, according to which birds in reduced condition would be more susceptible to IAV infection. The mechanisms responsible for reducing host competency among birds in poor condition remain unknown. Our research indicates body condition may influence the maintenance and spread of LPAIV by migrating waterfowl. ?? 2011 Arsnoe et al.
Arsnoe, Dustin M.; Ip, Hon S.; Owen, Jennifer C.
Migrating waterfowl are implicated in the global spread of influenza A viruses (IAVs), and mallards (Anas platyrhynchos) are considered a particularly important IAV reservoir. Prevalence of IAV infection in waterfowl peaks during autumn pre-migration staging and then declines as birds reach wintering areas. Migration is energetically costly and birds often experience declines in body condition that may suppress immune function. We assessed how body condition affects susceptibility to infection, viral shedding and antibody production in wild-caught and captive-bred juvenile mallards challenged with low pathogenic avian influenza virus (LPAIV) H5N9. Wild mallards (n = 30) were separated into three experimental groups; each manipulated through food availability to a different condition level (-20%, -10%, and normal ±5% original body condition), and captive-bred mallards (n = 10) were maintained at normal condition. We found that wild mallards in normal condition were more susceptible to LPAIV infection, shed higher peak viral loads and shed viral RNA more frequently compared to birds in poor condition. Antibody production did not differ according to condition. We found that wild mallards did not differ from captive-bred mallards in viral intensity and duration of infection, but they did exhibit lower antibody titers and greater variation in viral load. Our findings suggest that reduced body condition negatively influences waterfowl host competence to LPAIV infection. This observation is contradictory to the recently proposed condition-dependent hypothesis, according to which birds in reduced condition would be more susceptible to IAV infection. The mechanisms responsible for reducing host competency among birds in poor condition remain unknown. Our research indicates body condition may influence the maintenance and spread of LPAIV by migrating waterfowl.
Andrada, M; Quesada-Canales, O; Suárez-Bonnet, A; Paz-Sánchez, Y; Espinosa de Los Monteros, A; Rodríguez, F
Porcine enzootic pneumonia, primarily caused by Mycoplasma hyopneumoniae (Mh), is a contagious disease characterized by catarrhal bronchointerstitial pneumonia. Previous studies have evaluated immunohistochemically the distribution of Mh, different cellular populations and cytokines during Mh-induced pneumonia. Cyclooxygenase (COX)-2 is overexpressed during inflammatory responses by different cell types in the lung. The aim of this study was to elucidate the possible role of COX-2 in the pathogenesis of porcine enzootic pneumonia. COX-2 protein was detected by immunohistochemistry in formalin-fixed, paraffin wax-embedded lung tissues from 10 pigs infected experimentally with Mh. Ten pigs were inoculated intranasally with Mh and killed in pairs weekly from 1 to 5 weeks post inoculation. Three Mh-free pigs were taken as controls. Bronchial and bronchiolar epithelial cells, bronchial submucosal glands and a small number of macrophages in the bronchoalveolar exudate expressed COX-2. COX-2 protein was always associated with areas of pneumonia and expression was minimal in lungs from control pigs. These results suggest that COX-2 plays a role in the pathogenesis of Mh-infection.
Gowen, Brian B; Bray, Mike
A number of viruses in the family Arenaviridae cause severe illness in humans. Lassa virus in West Africa and a number of agents in South America produce hemorrhagic fever in persons exposed to aerosolized excretions of the pathogens' rodent hosts. Because arenaviruses are not transmitted by arthropods, and person-to-person spread is rare, human infections occur singly and sporadically, and are usually not diagnosed until the patient is severely ill. Because the arenaviruses are naturally transmitted by the airborne route, they also pose a potential threat as aerosolized bioterror weapons. The broad-spectrum antiviral drug ribavirin was shown to reduce mortality from Lassa fever, and has been tested against Argentine hemorrhagic fever, but it is not an approved treatment for either disease. Human immune convalescent plasma was proven to be effective for Argentine hemorrhagic fever in a controlled trial. New treatments are needed to block viral replication without causing toxicity and to prevent the increased vascular permeability that is responsible for hypotension and shock. In this paper, we review current developments in the experimental therapy of severe arenaviral infections, focusing on drugs that have been tested in animal models, and provide a perspective on future research.
Gowen, Brian B.; Bray, Mike
Summary A number of viruses in the family Arenaviridae cause severe illness in humans. Lassa virus in West Africa and a number of agents in South America produce hemorrhagic fever (HF) in persons exposed to aerosolized excretions of the pathogens’ rodent hosts. Because arenaviruses are not transmitted by arthropods, and person-to-person spread is rare, human infections occur singly and sporadically, and are usually not diagnosed until the patient is severely ill. Because the arenaviruses are naturally transmitted by the airborne route, they also pose a potential threat as aerosolized bioterror weapons. The broad-spectrum antiviral drug ribavirin was shown to reduce mortality from Lassa fever, and has been tested against Argentine HF, but it is not an approved treatment for either disease. Human immune convalescent plasma was proven to be effective for Argentine HF in a controlled trial. New treatments are needed to block viral replication without causing toxicity and to prevent the increased vascular permeability that is responsible for hypotension and shock. In this paper, we review current developments in the experimental therapy of severe arenaviral infections, focusing on drugs that have been tested in animal models, and provide a perspective on future research. PMID:22122440
Chen, P B; Davern, L B; Schifferle, R; Zambon, J J
The effects of immunization in modulating the pathogenesis of Bacteroides (Porphyromonas) gingivalis infection in a murine model system were examined. BALB/c mice were immunized by intraperitoneal injection with B. gingivalis ATCC 53977 (one injection per week for 3 weeks), or with a lithium diiodosalicylate (LIS) extract (one injection per week for 3 weeks), or with lipopolysaccharide (LPS; one intravenous or intraperitoneal injection) from this same strain. Two weeks after the final immunization, the mice were challenged by subcutaneous injection of B. gingivalis ATCC 53977. Mice immunized with bacteria had no secondary lesions and no septicemia, whereas mice immunized with LIS extract had few secondary lesions and no septicemia. Mice immunized with LPS and nonimmunized mice demonstrated secondary abdominal lesions and septicemia after challenge. Bacterial cells and LIS extract, but not LPS, induced serum antibody and antigen reactive lymphocytes, as measured by enzyme-linked immunosorbent assay, immunoblot, Western immunoblot transfer, and in vitro lymphoproliferative responses. The present study suggests that immunization with a LIS extract or whole cells may induce a protective response against experimental B. gingivalis infection. Images PMID:2401568
Maeda, Hiroki; Hatta, Takeshi; Alim, M Abdul; Tsubokawa, Daigo; Mikami, Fusako; Matsubayashi, Makoto; Miyoshi, Takeharu; Umemiya-Shirafuji, Rika; Kawazu, Shin-ichiro; Igarashi, Ikuo; Mochizuki, Masami; Tsuji, Naotoshi; Tanaka, Tetsuya
Ticks are potent vectors of many deadly human and animal pathogens. Tick-borne babesiosis is a well-recognized malaria-like disease that occurs worldwide and recently has attracted increased attention as an emerging zoonosis. Although the proliferation of Babesia organisms is essential in the vectors, their detailed lifecycle with time information for migration in ticks remains unknown. A novel study model for the elucidation of the migration speed of Babesia parasites in their vector tick, Haemaphysalis longicornis, has been developed using an artificial feeding system with quantitative PCR method. The detectable DNA of Babesia parasites gradually disappeared in the tick midgut at 1 day post engorgement (DPE), and in contrary increased in other organs. The results indicated that the Babesia parasite passed the H. longicornis midgut within 24 hours post engorgement, migrated to the hemolymph, and then proliferated in the organs except the midgut. This time point may be an important curfew for Babesia parasites to migrate in the tick lumen. We also visualized the Babesia parasites in the experimentally infected ticks and in their eggs using IFAT for detecting their cytoskeletal structure, which suggested the successful tick infection and transovarial transmission of the parasite. This model will shed light on the further understanding of tick-Babesia interactions. PMID:27841321
Chinnadurai, Sathya K; Cooper, David; Dombrowski, Daniel S; Poore, Matthew F; Levy, Michael G
Chytridiomycosis is an often fatal fungal disease of amphibians caused by Batrachochytrium dendrobatidis. This disease has been implicated in the worldwide decline of many anuran species, but studies of chytridiomycosis in wild salamanders are limited. Between August 2006 and December 2006, we tested wild amphibians in North Carolina, USA (n=212) by polymerase chain reaction (PCR). We identified three PCR-positive animals: one Rana clamitans and two Plethodontid salamanders. We experimentally infected two species of native North Carolina Plethodontid salamanders, the slimy salamander (Plethodon glutinosus) and the Blue Ridge Mountain dusky salamander (Desmognathus orestes) with 1,000,000 zoospores of B. dendrobatidis per animal. Susceptibility was species dependent; all slimy salamanders developed clinical signs of chytridiomycosis, and one died, whereas dusky salamanders remained unaffected. In a second experiment, we challenged naïve slimy salamanders with either 10,000 or 100,000 motile zoospores per animal. Clinical signs consistent with chytridiomycosis were not observed at either dose or in uninfected controls during the 45 days of this experiment. All animals inoculated with B. dendrobatidis in both experiments, regardless of dose, tested positive by PCR. Our study indicates that slimy salamanders are more susceptible to clinical chytridiomycosis than dusky salamanders, and in a laboratory setting, a dose greater than 100,000 zoospores per animal is required to induce clinical disease. This study also indicates that PCR is a very sensitive tool for detecting B. dendrobatidis infection, even in animals that are clinically unaffected, thus positive results should be interpreted with caution.
Stolle, A; Sperner, B
The goal of this presentation was to clarify which foods are involved in viral diseases, which viruses are transmitted via food and how to evaluate the risk of a foodborne viral infection. Food items frequently identified as cause of viral disease outbreaks were shellfish harvested in sewage-contaminated water. Another common source of foodborne viral illness was cold food contaminated by infected food handlers. In the European Union the viruses most frequently associated with foodborne illness were hepatitis A virus and the SRSV's. A few isolated cases of foodborne hepatitis E were reported in Mediterranean countries. Compared to other foodborne diseases, those caused by viruses are less severe and seldom fatal. This might be a reason why the problem of viral contamination of food has been neglected. Yet, because many foodborne viral diseases are not recognized either as foodborne or as caused by viruses, the actual number of cases must be assumed to be significantly higher than the reported number. Consequently, food associated diseases of viral origin should be granted more attention.
Shepherd, A. J.; Leman, P. A.; Hummitzsch, D. E.
Susceptibility studies were undertaken to determine the response of some South African wild rodent species to experimental plague (Yersinia pestis) infection. A degree of plague resistance was found in three gerbil species captured in the plague enzootic region of the northern Cape Province, these being the Namaqua gerbil, Desmodillus auricularis, (LD50 1 X 10(6) organisms), the bushveld gerbil, Tatera leucogaster, (LD50 9.1 X 10(5)) and the highveld gerbil, T. brantsii (LD50 4 X 10(2)). Animals from a population of the four-striped mouse, Rhabdomys pumilio, captured in the plague area of Port Elizabeth, proved moderately resistant to experimental plague infection (LD50 1.3 X 10(4)) while those from another population of the same species captured in a plague-free area of the Orange Free State were extremely susceptible (LD50, 5 organisms). The response of both populations however was a heterogeneous one. Marked differences in susceptibility were also found between two populations of multimammate mice, Mastomys natalensis (2n = 32) although both originated from areas outwith the known distribution of plague in southern Africa. The 50% infectious dose was relatively high in T. leucogaster (3.2 X 10(2)) and D. auricularis (1.7 X 10(3)), but was low (2-16 organisms) in the other rodent species tested. The plague antibody response, determined by enzyme-linked immunosorbent assay (ELISA), was extremely short-lived in T. leucogaster, only 10% of inoculated animals remaining seropositive at low titres after 11 weeks. Antibodies persisted for only slightly longer in the sera of T. brantsii which were reinoculated with 2 X 10(3) plague organisms 6 weeks after initial challenge. The demonstration of the existence of both susceptible and resistant populations of R. pumilio and M. natalensis indicates that these species must be considered as potential plague reservoir hosts in parts of South Africa. The results suggest that resistance to plague infection in previously epizootic
Canavarro, Maria C; Pereira, Marco; Simoes, Mario R; Pintassilgo, Ana L
The assessment of quality of life (QOL) in HIV infection has emerged as being vital to research and clinical practice. This assessment is also a challenge due to the specific characteristics of the infection, the increased availability of therapeutics, as well as the epidemiological variability inherent to HIV infection. The purpose of this study was to investigate the psychometric properties of the European Portuguese version of the World Health Organization's QOL Instrument in HIV Infection (WHOQOL-HIV) and to test its performance in a sample of HIV-infected patients. The European Portuguese version of WHOQOL-HIV was administered in a sample of 200 HIV-positive patients. The patients also completed the Portuguese versions of Beck Depression Inventory (BDI) and Brief Symptom Inventory (BSI). The WHOQOL-HIV showed quite an acceptable internal consistency (Cronbach's α ranged from 0.86 to 0.95 across domains). Convergent validity with BDI and BSI was satisfactory for all domains (all r>0.50; p<0.001). Moreover, correlations between domains and between domains and overall QOL were all statistically significant (p<0.001). The reliability and validity studies of the European Portuguese version of the WHOQOL-HIV revealed good psychometric characteristics, which allows for the use of this version of WHOQOL in our country, and cross-cultural comparability.
Bertelsen, Mads F; Meyland-Smith, Frederik; Willesen, Jakob L; Jefferies, Ryan; Morgan, Eric R; Monrad, Jesper
Metastrongyloid induced pneumonia has been described sporadically in the red panda (Ailurus fulgens). Early descriptions in pandas recently imported to the USA from China involved parasites morphologically similar to Angiostrongylus spp. and Crenosomatidae. More recently, four cases of severe verminous pneumonia associated with Angiostrongylus vasorum have been reported from European zoos. A coprological survey of the red panda population within European zoos was conducted in 2008. Faecal samples from 115 pandas originating from 54 zoos were collected on 3 consecutive days. Using Baermann technique, 40 animals (35%) from 20 zoos (37%) were found to shed metastrongyloid first stage larvae (L(1)). Based on their morphology and size, the L(1) observed could be divided into three morphologically distinct types: (1) a Crenosoma sp. type (n=5, overall prevalence: 4.3%), (2) an A. vasorum type (n=3, 2.6%), and (3) an unidentified metastrongyloid species, similar to, but morphologically distinct from A. vasorum (n=32, 27.8%). Further confirmation of species identification was provided by PCR amplification and sequencing of the 18S rRNA gene, which confirmed three different species. The novel Crenosoma species was most genetically analogous to Crenosoma mephitidis and the unidentified metastrongyloid species was most similar to Stenurus minor and Torynurus convulutus. Routine and quarantine health care of red pandas in captivity should take account of the risk of Angiostrongylus and Crenosoma infection in endemic areas, but should also be cognisant of the widespread presence of an apparently less pathogenic species of lungworm. The identity of the two potentially novel species is subject to further work.
Desmarets, Lowiese M. B.; Vermeulen, Ben L.; Theuns, Sebastiaan; Conceição-Neto, Nádia; Zeller, Mark; Roukaerts, Inge D. M.; Acar, Delphine D.; Olyslaegers, Dominique A. J.; Van Ranst, Marc; Matthijnssens, Jelle; Nauwynck, Hans J.
Feline infectious peritonitis (FIP) results from mutations in the viral genome during a common feline enteric coronavirus (FECV) infection. Since many virological and immunological data on FECV infections are lacking, the present study investigated these missing links during experimental infection of three SPF cats with FECV strain UCD. Two cats showed mild clinical signs, faecal shedding of infectious virus from 4 dpi, a cell-associated viraemia at inconsistent time points from 5 dpi, a highly neutralising antibody response from 9 dpi, and no major abnormalities in leukocyte numbers. Faecal shedding lasted for 28–56 days, but virus shed during this stage was less infectious in enterocyte cultures and affected by mutations. Remarkably, in the other cat neither clinical signs nor acute shedding were seen, but virus was detected in blood cells from 3 dpi, and shedding of non-enterotropic, mutated viruses suddenly occurred from 14 dpi onwards. Neutralising antibodies arose from 21 dpi. Leukocyte numbers were not different compared to the other cats, except for the CD8+ regulatory T cells. These data indicate that FECV can infect immune cells even in the absence of intestinal replication and raise the hypothesis that the gradual adaptation to these cells can allow non-enterotropic mutants to arise. PMID:26822958
Rodríguez-Cortés, Alhelí; Ojeda, Ana; Todolí, Felicitat; Alberola, Jordi
Leishmania infantum (syn. Leishmania chagasi) is the etiological agent of a widespread serious zoonotic disease that affects both humans and dogs. Prevalence and incidence of the canine infection are important parameters to determine the risk and the ways to control this reemergent zoonosis. Unfortunately, there is not a gold standard test for Leishmania infection. Our aim was to assess the operative validity of commercial tests used to detect antibodies to Leishmania in serum samples from experimental infections. Three ELISA tests (LEISCAN(®) Leishmania ELISA Test, INGEZIM(®) LEISHMANIA, and INGEZIM(®) LEISHMANIA VET), three immunochromatographic tests (INGEZIM(®) LEISHMACROM, SNAP(®) Leishmania, and WITNESS(®) Leishmania), and one IFAT were evaluated. LEISCAN(®) Leishmania ELISA test achieved the highest sensitivity and accuracy (both 0.98). Specificity was 1 for all tests except for IFAT. All tests but IFAT obtained a positive predictive value of 1, while the maximum negative predictive value was achieved by LEISCAN(®) Leishmania ELISA Test (0.93). The best positive likelihood ratio was obtained by INGEZIM(®) LEISHMANIA VET (30.26), while the best negative likelihood ratio was obtained by LEISCAN(®) Leishmania ELISA Test (0.02). The highest diagnostic odds ratio was achieved by LEISCAN(®) Leishmania ELISA Test (729.00). The largest area under the ROC curve was obtained by LEISCAN(®) Leishmania ELISA Test (0.981). Quantitative ELISA based tests performmed better than qualitative tests ("Rapid Tests"), and the test best suited to detect Leishmania in infected dogs and to provide clinically useful information was LEISCAN(®) Leishmania ELISA Test. This and other results point also to the need of revising the status of IFAT as a gold standard for the diagnosis of leishmaniasis.
Ahmed, S T; Mun, H-S; Yoe, H; Yang, C-J
Behavior is one of the most commonly used indicators of illness; however, few studies have investigated how different common diseases affect animal behavior. This experiment was conducted to investigate behavioral and clinical alterations in growing pigs experimentally infected with Salmonella spp. during a 4-week post-infection period. A total of 48 growing pigs were divided into one of the three treatment groups (1) control, (2) infection with Salmonella Typhimurium or (3) infection with Salmonella Enteritidis. Individual pigs' behavior was recorded daily (0900 to 1100 and 1600 to 1800 h) using a video-recording system. Pigs in both infected groups had lower weight gain and feed intake during week 0 to 2 and 0 to 4 experimental period. Bacteriological data revealed that pigs in both infected groups persistently shed bacteria throughout the period of study. Oral infection of growing pigs with S. Typhimurium and S. Enteritidis significantly reduced the frequency of morning large (except week 1) and small movement throughout the study period. In the evening, significantly lowest frequency of movements were observed in the S. Enteritidis-infected group compared with the control. The standing and sitting frequency were significantly lower in both infected groups only at the morning of week 4. Infection with Salmonella spp. led to a significant reduction in the frequency and duration of morning eating and drinking throughout the experimental period, with the exception of 4th week drinking duration. The lowest frequency of evening eating during week 1 and 4 was recorded in both infected groups; whereas, the duration differed only at week 1. The evening drinking frequency only tended to decrease in response to S. Typhimurium infection at week 1. This study shows that, pigs infected with Salmonella spp. had poor performance, shedding high levels of Salmonella with their feces and reduced feeding and drinking activity, which are adaptive responses to infection and may help
Pozio, E; Casulli, A; Bologov, V V; Marucci, G; La Rosa, G
From 1998 to 2000, 184 animals (82 wolves, 29 red foxes, 55 mustelids, 5 raccoon dogs, and 13 domestic dogs), mainly shot by hunters in the Tvier and Smoliensk regions of northwest European Russia, were tested for Trichinella larvae; 98 animals (53.3%) were found to be positive. The highest prevalence was detected in wolf (97.5%). Trichinella nativa was the most common species detected (98%). The diet of wolves was investigated by examining the stomach contents of 62 animals (75.6% of the total number of wolves examined for Trichinella). It consisted mainly of dog (36.4% of the total number of occurrences of all food items, PFO) and moose (31.2 PFO); however, during the hunting seasons of 1998-1999 and 1999-2000, skinned wolf carcasses were left in the forest as bait (567 carcasses, about 18,000 kg). This very high prevalence of Trichinella infection, the highest ever detected in a natural population of carnivores, could be explained by carnivore-carnivore transmission, influenced by the hunting practices adopted in the study area.
Pes, Giovanni Mario; Tognotti, Eugenia; Poulain, Michel; Chambre, Dany; Dore, Maria Pina
Since ancient times the Mediterranean island of Sardinia has been known for harboring a population with an average body height shorter than almost every other ethnic group in Europe. After over a century of investigations, the cause(s) at the origin of this uniqueness are not yet clear. The shorter stature of Sardinians appears to have been documented since prehistoric times, as revealed by the analysis of skeletal remains discovered in archaeological sites on the island. Recently, a number of genetic, hormonal, environmental, infective and nutritional factors have been put forward to explain this unique anthropometric feature, which persisted for a long time, even when environmental and living conditions improved around 1960. Although some of the putative factors are supported by sound empirical evidence, weaker support is available for others. The recent advent of whole genome analysis techniques shed new light on specific variants at the origin of this short stature. However, the marked geographical variability of stature across time and space within the island, and the well-known presence of pockets of short height in the population of the southern districts, are still puzzling findings that have attracted the interest of anthropologists and geneticists. The purpose of this review is to focus on the state-of-the-art research on stature, as well as the factors that made Sardinians the shortest among Europeans.
Brijs, Jeroen; Jutfelt, Fredrik; Clark, Timothy D; Gräns, Albin; Ekström, Andreas; Sandblom, Erik
A progressive inability of the cardiorespiratory system to maintain systemic oxygen supply at elevated temperatures has been suggested to reduce aerobic scope and the upper thermal limit of aquatic ectotherms. However, few studies have directly investigated the dependence of thermal limits on oxygen transport capacity. By manipulating oxygen availability (via environmental hyperoxia) and blood oxygen carrying capacity (via experimentally induced anaemia) in European perch (Perca fluviatilis Linneaus), we investigated the effects of oxygen transport capacity on aerobic scope and the critical thermal maximum (CT(max)). Hyperoxia resulted in a twofold increase in aerobic scope at the control temperature of 23°C, but this did not translate to an elevated CT(max) in comparison with control fish (34.6±0.1 versus 34.0±0.5°C, respectively). Anaemia (∼43% reduction in haemoglobin concentration) did not cause a reduction in aerobic scope or CT(max) (33.8±0.3°C) compared with control fish. Additionally, oxygen consumption rates of anaemic perch during thermal ramping increased in a similar exponential manner to that in control fish, highlighting that perch have an impressive capacity to compensate for a substantial reduction in blood oxygen carrying capacity. Taken together, these results indicate that oxygen limitation is not a universal mechanism determining the CT(max) of fishes.
Uhrich, Pierre J. M.; Juompan, B.; Tourde, R.; Brunet, M.; Dutrey, J.-F.
The CNES project of a European Complement to GPS (CE-GPS) is conceived to fulfill the needs of Civil Aviation for a non-precise approach phase with GPS as sole navigation means. This generates two missions: a monitoring mission - alarm of failure - ,and a navigation mission - generating a GPS-like signal on board the geostationary satellites. The host satellites will be the Inmarsat constellation. The CE-GPS missions lead to some time requirements, mainly the accuracy of GPS time restitution and of monitoring clock synchronization. To demonstrate that the requirements of the CE-GPS could be achieved, including the time aspects, an experiment has been scheduled over the Last two years, using a part of the Inmarsat II F-2 payload and specially designed ground stations based on 10 channels GPS receivers. This paper presents a review of the results obtained during the continental phase of the CE-GPS experiment with two stations in France, along with some experimental results obtained during the transatlantic phase (three stations in France, French Guyana, and South Africa). It describes the synchronization of the monitoring clocks using the GPS Common-view or the C- to L-Band transponder of the Inmarsat satellite, with an estimated accuracy better than 10 ns (1 sigma).
Stone, Robert J.
Space Telerobotics research, performed under contract to the European Space Agency (ESA), concerning the execution of human factors experiments, and ultimately leading to the development of a telerobotics test bed, has been carried out since 1985 by a British Consortium consisting of British Aerospace, the United Kingdom Atomic Energy Authority and, more recently, the UK National Advanced Robotics Research Centre. The principal aim of the first study of the series was to derive preliminary requirements for a teleoperation servicing system, with reference to two mission model scenarios. The first scenario introduced the problem of communications time delays, and their likely effect on the ground-based operator in control of a manipulator system on board an unmanned servicing vehicle in Low Earth Orbit. In the second scenario, the operator was located on the NASA Orbiter aft flight deck, supervising the control of a prototype manipulator in the 'servicing' of an experimental payload in the cargo bay area. Human factors analyses centered on defining the requirements for the teleoperator workstation, such as identifying basic ergonomic requirements for workstation and panel layouts, defining teleoperation strategies, developing alphanumeric and graphic screen formats for the supervision or direct control of the manipulator, and the potential applications of expert system technology. The second study for ESA involved an experimental appraisal of some of the important issues highlighted in the first study, for which relevant human factors data did not exist. Of central importance during the second study was the issue of communications time delays and their effect on the manual control of a teleoperated manipulator from a ground-based command and control station.
Averbuch, Diana; Orasch, Christina; Cordonnier, Catherine; Livermore, David M; Mikulska, Malgorzata; Viscoli, Claudio; Gyssens, Inge C; Kern, Winfried V; Klyasova, Galina; Marchetti, Oscar; Engelhard, Dan; Akova, Murat
Owing to increasing resistance and the limited arsenal of new antibiotics, especially against Gram-negative pathogens, carefully designed antibiotic regimens are obligatory for febrile neutropenic patients, along with effective infection control. The Expert Group of the 4(th) European Conference on Infections in Leukemia has developed guidelines for initial empirical therapy in febrile neutropenic patients, based on: i) the local resistance epidemiology; and ii) the patient's risk factors for resistant bacteria and for a complicated clinical course. An 'escalation' approach, avoiding empirical carbapenems and combinations, should be employed in patients without particular risk factors. A 'de-escalation' approach, with initial broad-spectrum antibiotics or combinations, should be used only in those patients with: i) known prior colonization or infection with resistant pathogens; or ii) complicated presentation; or iii) in centers where resistant pathogens are prevalent at the onset of febrile neutropenia. In the latter case, infection control and antibiotic stewardship also need urgent review. Modification of the initial regimen at 72-96 h should be based on the patient's clinical course and the microbiological results. Discontinuation of antibiotics after 72 h or later should be considered in neutropenic patients with fever of unknown origin who are hemodynamically stable since presentation and afebrile for at least 48 h, irrespective of neutrophil count and expected duration of neutropenia. This strategy aims to minimize the collateral damage associated with antibiotic overuse, and the further selection of resistance.
Averbuch, Diana; Orasch, Christina; Cordonnier, Catherine; Livermore, David M.; Mikulska, Małgorzata; Viscoli, Claudio; Gyssens, Inge C.; Kern, Winfried V.; Klyasova, Galina; Marchetti, Oscar; Engelhard, Dan; Akova, Murat
Owing to increasing resistance and the limited arsenal of new antibiotics, especially against Gram-negative pathogens, carefully designed antibiotic regimens are obligatory for febrile neutropenic patients, along with effective infection control. The Expert Group of the 4th European Conference on Infections in Leukemia has developed guidelines for initial empirical therapy in febrile neutropenic patients, based on: i) the local resistance epidemiology; and ii) the patient’s risk factors for resistant bacteria and for a complicated clinical course. An ‘escalation’ approach, avoiding empirical carbapenems and combinations, should be employed in patients without particular risk factors. A ‘de-escalation’ approach, with initial broad-spectrum antibiotics or combinations, should be used only in those patients with: i) known prior colonization or infection with resistant pathogens; or ii) complicated presentation; or iii) in centers where resistant pathogens are prevalent at the onset of febrile neutropenia. In the latter case, infection control and antibiotic stewardship also need urgent review. Modification of the initial regimen at 72–96 h should be based on the patient’s clinical course and the microbiological results. Discontinuation of antibiotics after 72 h or later should be considered in neutropenic patients with fever of unknown origin who are hemodynamically stable since presentation and afebrile for at least 48 h, irrespective of neutrophil count and expected duration of neutropenia. This strategy aims to minimize the collateral damage associated with antibiotic overuse, and the further selection of resistance. PMID:24323983
King, Tracey Anne; van Oosterhout, Cock; Cable, Joanne
Introduced exotic species have the potential to spread their associated parasites to native species which can be catastrophic if these hosts are immunologically naïve to the novel parasite. The guppy (Poecilia reticulata) has been disseminated worldwide outside of its native habitat and therefore could be an important source of infection to native fish species. Its parasite fauna is dominated by the ectoparasitic monogeneans, Gyrodactylus turnbulli and Gyrodactylus bullatarudis. The current study tested the host specificity of G. bullatarudis by experimentally infecting a range of isolated fish hosts, including temperate species. Surprisingly, the parasite was capable of establishing and reproducing, for several days, on the three-spined stickleback when transferred directly to this host. We also established that G. bullatarudis could be transmitted under aquarium conditions at both 25 degrees C and 15 degrees C. At the higher temperature, the parasite was even capable of reproducing on this atypical host. The implications of these findings are discussed in terms of host specificity, host switching and climate change.
Kocan, Katherine M; Busby, Ann T; Allison, Robin W; Breshears, Melanie A; Coburn, Lisa; Galindo, Ruth C; Ayllón, Nieves; Blouin, Edmour F; de la Fuente, José
Anaplasma phagocytophilum, first identified as a pathogen of ruminants in Europe, has more recently been recognized as an emerging tick-borne pathogen of humans in the U.S. and Europe. A. phagocytophilum is transmitted by Ixodes spp., but the tick developmental cycle and pathogen/vector interactions have not been fully described. In this research, we report on the experimental infection of sheep with the human NY-18 isolate of A. phagocytophilum which then served as a host for infection of I. scapularis nymphs and adults. A. phagocytophilum was propagated in the human promyelocytic cell line, HL-60, and the infected cell cultures were then used to infect sheep by intravenous inoculation. Infections in sheep were confirmed by PCR and an Anaplasma-competitive ELISA. Clinical signs were not apparent in any of the infected sheep, and only limited hematologic and mild serum biochemical abnormalities were identified. While A. phagocytophilum morulae were rarely seen in neutrophils, blood film evaluation revealed prominent large granular lymphocytes, occasional plasma cells, and rare macrophages. Upon necropsy, gross lesions were restricted to the lymphoid system. Mild splenomegaly and lymphadenomegaly with microscopic evidence of lymphoid hyperplasia was observed in all infected sheep. Female I. scapularis that were allowed to feed and acquire infection on each of the 3 experimentally infected sheep became infected with A. phagocytophilum as determined by PCR of guts (80-87%) and salivary glands (67-100%). Female I. scapularis that acquired infection as nymphs on an experimentally infected sheep transmitted A. phagocytophilum to a susceptible sheep, thus confirming transstadial transmission. Sheep proved to be a good host for the production of I. scapularis infected with this human isolate of A. phagocytophilum, which can be used as a model for future studies of the tick/pathogen interface.
Popielarczyk, R; Robak, S; Siwicki, K A
The aim of this study was to determine the degree of Anguillicoloides crassus infection in European eel inhabiting Polish waters based on selected parasitic descriptors and on anatomical pathology of the swimbladder using macroscopic methods. In all, 154 European eel specimens were sampled from eleven sites in Poland and A. crassus was present in the swimbladder of 114 fish. The intensity of A. crassus infection in all the eel specimens ranged from 1 to 62 parasites at a mean value of 7.5. High values of mean infection intensity were noted in samples from Pomeranian lakes Bukowo, Łebsko, and Jamno. The health of the swimbladder was evaluated using the swimbladder degenerative index (SDI). The mean value of the SDI for all of the eel examined was 3.3, and extensively degenerated swimbladders were observed mainly in samples in the Szczecin Lagoon and from lakes. According to the individual SDI ratings, 9.1% of the eel specimens did not exhibit pathological symptoms of the swimbladder (SDI-0) and an extremely damaged (SDI-6) swimbladder was noted in 11.7% of the fish examined. In the case of eel infected with A. crassus, higher SDI values were reflected in initially increasing shares in subsequent categories. In fish that were not infected with the nematode, only 20% (8 individuals) of the swimbladders showed no symptoms of pathology (SDI-0).
Enderlein, Dirk; Herzog, Sibylle; Kaleta, Erhard F.; Heffels-Redmann, Ursula; Ressmeyer, Saskia; Herden, Christiane; Lierz, Michael
Avian bornavirus (ABV) is the presumed causative agent of proventricular dilatation disease (PDD), a major fatal disease in psittacines. However, the influencing factors and pathogenesis of PDD are not known and natural ABV infection exhibits remarkable variability. We investigated the course of infection in 18 cockatiels that were intracerebrally and intravenously inoculated with ABV. A persistent ABV infection developed in all 18 cockatiels, but, as in natural infection, clinical disease patterns varied. Over 33 weeks, we simultaneously studied seroconversion, presence of viral RNA and antigens, infectious virus, histopathologic alterations, and clinical signs of infection in the ABV-infected birds. Our study results further confirm the etiologic role of ABV in the development of PDD, and they provide basis for further investigations of the pathogenetic mechanisms and disease-inducing factors for the development of PDD. PMID:22304809
Delahay, R J; Cheeseman, C L; Clifton-Hadley, R S
Mycobacterium bovis infection has been confirmed in a wide range of mammals hosts throughout the world. The European badger (Meles meles) and the brushtail possum (Trichosurus vulpecula) are implicated as significant sources of infection for domestic cattle in the UK and New Zealand respectively. The risk of transmission of infection between a wildlife population and domestic animals will be determined by both the epidemiology of the disease and the ecology of the host. In the UK, surveys by the UK Ministry of Agriculture, Fisheries and Food (MAFF) have identified M. bovis infection in deer (Cervus sp., Capreolus sp., Dama sp.), red fox (Vulpes vulpes), mink (Mustela vison), feral ferret (Mustela furo), mole (Talpa europaea), brown rat (Rattus norvegicus) and feral cat (Felis catus). However, the potential contribution to cattle herd breakdowns, of reservoirs of M. bovis infection in mammals other than the badger is poorly understood and is the subject of current research. In contrast, M. bovis infection in the badger has been the subject of a long term ecological and epidemiological study at Woodchester Park in South-West England, where the prevalence and distribution of infection in a wild population has been intensively monitored. The pattern of infection in the population and potential risks to cattle, are profoundly influenced by badger social organization and behaviour. The pattern of land use and cattle farming practices in the UK brings badgers into close contact with domestic animals and provides conditions that may enhance the likelihood of disease transfer.
Tomlinson, A J; Chambers, M A; Wilson, G J; McDonald, R A; Delahay, R J
Heterogeneity in the progression of disease amongst individual wild animals may impact on both pathogen and host dynamics at the population level, through differential effects on transmission, mortality and reproductive output. The role of the European badger (Meles meles) as a reservoir host for Mycobacterium bovis infection in the UK and Ireland has been the focus of intense research for many years. Here, we investigate life-history correlates of infection in a high-density undisturbed badger population naturally infected with M. bovis. We found no evidence of a significant impact of M. bovis infection on female reproductive activity or success, with evidence of reproduction continuing successfully for several years in the face of M. bovis excretion. We also found evidence to support the hypothesis that female badgers are more resilient to established M. bovis infection than male badgers, with longer survival times following the detection of bacterial excretion. We discuss the importance of infectious breeding females in the persistence of M. bovis in badger populations, and how our findings in male badgers are consistent with testosterone-induced immunosuppression. In addition, we found significant weight loss in badgers with evidence of disseminated infection, based on the culture of M. bovis from body systems other than the respiratory tract. For females, there was a gradual loss of weight as infection progressed, whereas males only experienced substantial weight loss when infection had progressed to the point of dissemination. We discuss how these differences may be explained in terms of resource allocation and physiological trade-offs.
Deblanc, C; Gorin, S; Quéguiner, S; Gautier-Bouchardon, A V; Ferré, S; Amenna, N; Cariolet, R; Simon, G
Swine influenza virus (SIV) and Mycoplasma hyopneumoniae (Mhp) are widespread in farms and are major pathogens involved in the porcine respiratory disease complex (PRDC). The aim of this experiment was to compare the pathogenicity of European avian-like swine H1N1 and European human-like reassortant swine H1N2 viruses in naïve pigs and in pigs previously infected with Mhp. Six groups of SPF pigs were inoculated intra-tracheally with either Mhp, or H1N1, or H1N2 or Mhp+H1N1 or Mhp+H1N2, both pathogens being inoculated at 21 days intervals in these two last groups. A mock-infected group was included. Although both SIV strains induced clinical signs when singly inoculated, results indicated that the H1N2 SIV was more pathogenic than the H1N1 virus, with an earlier shedding and a greater spread in lungs. Initial infection with Mhp before SIV inoculation increased flu clinical signs and pathogenesis (hyperthermia, loss of appetite, pneumonia lesions) due to the H1N1 virus but did not modify significantly outcomes of H1N2 infection. Thus, Mhp and SIV H1N1 appeared to act synergistically, whereas Mhp and SIV H1N2 would compete, as H1N2 infection led to the elimination of Mhp in lung diaphragmatic lobes. In conclusion, SIV would be a risk factor for the severity of respiratory disorders when associated with Mhp, depending on the viral subtype involved. This experimental model of coinfection with Mhp and avian-like swine H1N1 is a relevant tool for studying the pathogenesis of SIV-associated PRDC and testing intervention strategies for the control of the disease.
Taira, Kensuke; Yanagida, Tomonori; Akazawa, Naoko; Saitoh, Yasuhide
The organ distribution of Toxocara cati larvae in albino rats Rattus norvegicus (n=6/group) experimentally inoculated with 1000 embryonated eggs was examined 1, 2, 3, 7, 30, 90, and 180 days post inoculation (dpi), and the infectivity of recovered larvae was evaluated by bioassay in mice. The intestines, liver, lungs, muscles (carcass) and other organs (heart, brain, spleen, kidneys and genital organs) were digested for larval recovery. Larvae were recovered from all rats, with the mean number of recovered larvae ranging from 13.3 at 1 dpi to 135.6 at 90 dpi. Most of the larvae recovered were detected in the intestines (56.3%) and liver (43.8%) at 1 dpi; liver (21.6%) and lungs (69.6%) at 2 dpi; muscles (45.9%) and lungs (36.9%) at 3 dpi. Subsequently, most of larvae were recovered from muscles at 7 dpi (92.5%), 30 dpi (97.8%), 90 dpi (99.4%) and 180 dpi (99.1%). In the mouse bioassay, 43.8% of 90-day-old larvae and 43.0% of 180-day-old larvae recovered from rats established in mice. The present study demonstrated that T. cati larvae persist predominantly in rat muscles and nearly half of them retain infective for at least half a year. The results indicate that R. norvegicus may be a suitable paratenic host of T. cati under natural conditions.
Montiel, Nestor; Buckley, Alexandra; Guo, Baoqing; Kulshreshtha, Vikas; VanGeelen, Albert; Hoang, Hai; Rademacher, Christopher; Yoon, Kyoung-Jin
Senecavirus A has been infrequently associated with vesicular disease in swine since 1988. However, clinical disease has not been reproduced after experimental infection with this virus. We report vesicular disease in 9-week-old pigs after Sencavirus A infection by the intranasal route under experimental conditions. PMID:27315363
Montiel, Nestor; Buckley, Alexandra; Guo, Baoqing; Kulshreshtha, Vikas; VanGeelen, Albert; Hoang, Hai; Rademacher, Christopher; Yoon, Kyoung-Jin; Lager, Kelly
Senecavirus A has been infrequently associated with vesicular disease in swine since 1988. However, clinical disease has not been reproduced after experimental infection with this virus. Here we report vesicular disease in 9-week-old pigs after Sencavirus A infection by the intranasal route under experimental conditions.
Ratti, F.; Gavira, J.; Thirkettle, A. C.; Erba, F.; Muylaert, J.-M.; Walpot, L.; Rembiasz, J. M.
The European Experimental Re-entry Test-bed EXPERT is developed by the European Space Agency (ESA) as part of its General Technological Research Program (GSTP). The aim of EXPERT is to improve the understanding of critical aero-thermodynamic phenomena associated with hypersonic re-entry flights. The EXPERT project provides an opportunity to the scientific community and industries throughout Europe to propose and perform experiments in order to obtain aero-thermodynamic data for the validation of numerical models and of ground to flight extrapolation methodologies. During the last years an intense activity has been performed at ESA in order to select the most suitable experiments, bring them to a mature design, manufacture the qualification model and qualify the experiments for flight. ESA staffs coordinated and supported the work of the principal investigators of the experiments from European institutions and industrial organizations in order to maximize the scientific output in compliance with the budget resources made available to the EXPERT project and the programmatic constraints. EXPERT is a re-entry capsule having the shape of a blunted cone. The front part consists of a nose made of ceramic material developed at DLR Stuttgart. No ablative material is implemented so as not to contaminate the specific measurements of Payloads on board. The ceramic nose hosts a set of experiments: the Flush Air Data System (FADS) developed by HTG aiming at collecting free flow data required for post flight analysis, the pyrometer PYREX developed at IRS in Stuttgart collecting data on the temperature and heat flux of the ceramic nose, and the IRS spectrometer RESPECT aiming at resolving the different species generated in the plasma region during re-entry. The sides of the blunted cone are protected by a metallic thermal protection system in which several experiments are located. Two Payloads developed by IRS and VKI are dedicated to the measurement of catalytic effects. One aims
Market pigs infected with Salmonella pose a significant food safety risk by carrying the pathogen into abattoirs. A study was conducted to determine the dynamic of Salmonella infection in market-weight pigs (220-240 lbs.). Pigs (n=24) were individually inoculated (intranasally; 108 cfu/mL) with Salm...
Rocke, Tonie E.; Yuill, Thomas M.; Amundson, Terry E.
The effects of Mycoplasma gallisepticum (MG) infections on egg production, fertility, and hatchability were studied in captive-reared wild turkeys (Meleagris gallopavo). Three groups of adult birds, each consisting of four hens and two toms, were exposed to MG by the respiratory route at the beginning of their breeding season. Fourteen control birds received sterile growth medium. Although no mortality of infected or control birds occurred, egg production during the first breeding season after infection was reduced. The mean number of eggs/hen/day produced by infected groups the first breeding season postexposure (PE) was significantly lower than the control value. The mean number of eggs produced daily by the same hens 1 yr later was unaffected by MG infection. The pecentage of fertile eggs produced by infected groups was slightly reduced in both the first and second breeding seasons PE. Hatchability of fertile eggs from infected hens was significantly lower than eggs from control hens. Productivity may be impaired if MG infections occur in free-ranging wild turkey populations.
Kraus, S J; Jacobs, N F; Chandler, F W; Arum, E S
Subcutaneous tissue cavities in mice and guinea pigs were infected with human isolates of Ureaplasma urealyticum and Mycoplasma hominis. The minimal infective dose for M. hominis was as low as less than 10 color-changing units (CCU) for mice and 10(2) CCU for guinea pigs. The minimal infective dose for U. urealyticum was as low as less than 10 CCU for mice and 10(4) CCU for guinea pigs. Mouse infections with either U. urealyticum or M. hominis persisted for 1 day to greater than 4 months. Guinea pigs remained infected for up to 4 weeks. Two M. hominis isolates were similar in their ability to infect subcutaneous tissue cavities but two U. urealyticum isolates varied in their ability to infect the cavities. The histopathology of the M. hominis and U. urealyticum infections was similar: an initial intense polymorphonuclear response with giant cells, followed in 4 weeks by histiocytes and giant cells with some plasma cells and lymphocytes. Images PMID:873611
Crémieux, A C; Carbon, C
Bone and joint infections are difficult to cure. The difficulty is related to the presence of bacteria adherent to foreign material in many cases and also to the limited activity of antibiotics in infected bones. Clinical trials are difficult to design because of the heterogeneity of the disease and the number of factors that could influence the therapeutic response. To control for these multiple variables, attempts have been made to develop reliable animal models of osteomyelitis and prosthetic joint infections that closely mimic the different infections seen in orthopedic surgery and that allow evaluation of the efficacy of surgical procedures as well as local or systemic antibiotic therapy. These models will continue to provide us information on the pathogenesis and management of such infections.
Hope, V D; Eramova, I; Capurro, D; Donoghoe, M C
Knowledge of hepatitis B and C prevalence, and numbers infected, are important for planning responses. Published HBsAg and anti-HCV prevalences for the 20 WHO European Region countries outside the EU/EFTA were extracted, to complement published data for the EU/EFTA. The general population prevalence of HBsAg (median 3·8%, mean 5·0%, seven countries) ranged from 1·3% (Ukraine) to 13% (Uzbekistan), and anti-HCV (median 2·3%, mean 3·8%, 10 countries) from 0·5% (Serbia, Tajikistan) to 13% (Uzbekistan). People who inject drugs had the highest prevalence of both infections (HBsAg: median 6·8%, mean 8·2%, 13 countries; anti-HCV: median 46%, mean 46%, 17 countries), and prevalence was also elevated in men who have sex with men and sex workers. Simple estimates indicated 13·3 million (1·8%) adults have HBsAg and 15·0 million (2·0%) HCV RNA in the WHO European Region; prevalences were higher outside the EU/EFTA countries. Efforts to prevent, diagnose, and treat these infections need to be maintained and improved. This article may not be reprinted or reused in any way in order to promote any commercial products or services.
Ortiz-Flores, Aaxin M.; Ledesma-Soto, Yadira; Calleja, Elsa A.; Rodríguez-Sosa, Miriam; Juárez, Imelda; Terrazas, Luis I.
It was previously reported by our group that infection with Taenia crassiceps reduces incidence and severity of inflammatory and autoimmune experimental diseases like type 1 diabetes and experimental autoimmune encephalomyelitis. In this research, we set out to study whether infection with T. crassiceps would affect the development of experimental rheumatoid arthritis (RA). We found that mice infected with the parasite and induced with experimental RA showed similar clinical scores as the noninfected experimental RA group; systemic cytokines were not affected while anti-CII Abs were higher in the infected group. Histological evaluation showed damage in both infected and noninfected experimental RA-induced groups and although some surface molecules such as PDL-2 and MR which are associated with immunomodulatory mechanisms were upregulated in the infected and RA-induced group as compared to the noninfected RA group, they did not exert any changes in the outcome of experimental RA. Thus, we determined that infection with T. crassiceps does not influence the outcome of experimental RA. PMID:23509709
Ortiz-Flores, Aaxin M; Ledesma-Soto, Yadira; Calleja, Elsa A; Rodríguez-Sosa, Miriam; Juárez, Imelda; Terrazas, Luis I
It was previously reported by our group that infection with Taenia crassiceps reduces incidence and severity of inflammatory and autoimmune experimental diseases like type 1 diabetes and experimental autoimmune encephalomyelitis. In this research, we set out to study whether infection with T. crassiceps would affect the development of experimental rheumatoid arthritis (RA). We found that mice infected with the parasite and induced with experimental RA showed similar clinical scores as the noninfected experimental RA group; systemic cytokines were not affected while anti-CII Abs were higher in the infected group. Histological evaluation showed damage in both infected and noninfected experimental RA-induced groups and although some surface molecules such as PDL-2 and MR which are associated with immunomodulatory mechanisms were upregulated in the infected and RA-induced group as compared to the noninfected RA group, they did not exert any changes in the outcome of experimental RA. Thus, we determined that infection with T. crassiceps does not influence the outcome of experimental RA.
Sellon, R K; Jordan, H L; Kennedy-Stoskopf, S; Tompkins, M B; Tompkins, W A
Postnatal transmission of feline immunodeficiency virus (FIV) in neonates nursed by acutely infected mothers and infection resulting from oral inoculation of kittens with FIV were evaluated. Ten of 16 kittens nursed by four queens with FIV infection established immediately postpartum developed FIV infection. Five of 11 neonates orally administered cell-free FIV culture supernatant developed FIV infection. Kittens that developed FIV infection had greater proportions of CD4+ and Pan-T+ lymphocytes at birth than negative kittens. Infectious virus was recovered from the milk of acutely infected mothers. We conclude that FIV may be experimentally transmitted via milk from queens with acute infections and that oral administration of FIV to neonatal kittens results in infection. Images PMID:8151797
Styczynski, J; Reusser, P; Einsele, H; de la Camara, R; Cordonnier, C; Ward, K N; Ljungman, P; Engelhard, D
These guidelines on the management of HSV, VZV and EBV infection in patients with hematological malignancies and after SCT were prepared by the European Conference on Infections in Leukemia following a predefined methodology. A PubMed search was conducted using the appropriate key words to identify studies pertinent to management of HSV, VZV and EBV infections. References of relevant articles and abstracts from recent hematology and SCT scientific meetings were also reviewed. Prospective and retrospective studies identified from the data sources were evaluated, and all data deemed relevant were included in this analysis. The clinical and scientific background was described and discussed, and the quality of evidence and level of recommendation were graded according to the Centers for Disease Control criteria.
Doffitt, Cynthia M; Pote, Linda M; King, D Tommy
In order to determine potential definitive hosts of the digenetic trematode, Bolbophorus damnificus, two American White Pelicans (Pelecanus erythrorhynchos), two Double-crested Cormorants (Phalacrocorax auritus), two Great Blue Herons (Ardea herodias), and two Great Egrets (Ardea alba) were captured, treated with praziquantel, and fed channel catfish (Ictalurus punctatus) infected with B. damnificus metacercariae. Patent infections of B. damnificus, which developed in both American White Pelicans at 3 days postinfection, were confirmed by the presence of trematode ova in the feces. Mature B. damnificus trematodes were recovered from the intestines of both pelicans at 21 days postinfection, further confirming the establishment of infection. No evidence of B. damnificus infections was observed in the other bird species studied. This study provides further evidence that Double-crested Cormorants, Great Blue Herons, and Great Egrets do not serve as definitive hosts for B. damnificus.
Seong, Giyong; Oem, Jae-Ku; Lee, Kyung-Hyun; Choi, Kyoung-Seong
The objective of this study was to test the ability of bovine viral diarrhea virus (BVDV) to infect mice. Two mice each were either mock infected or inoculated with one of three BVDV strains by the intraperitoneal (IP) (n = 8) or intranasal (IN) (n = 8) route. All mice were euthanized at day 7 postinfection (p.i.). None of the infected mice exhibited any clinical signs of illness; however, the tissues harvested after BVDV challenge showed significant histopathological changes. Blood samples from five mice that were injected IP and one mouse that was inoculated IN were positive for BVDV by reverse transcription polymerase chain reaction (RT-PCR). Immunohistochemistry (IHC) was used to assess the presence of viral antigen in the organs of mice infected with three BVDV strains. In IP-injected mice, BVDV antigen was detected in the spleen (5/6), mesenteric lymph nodes (4/6), lymphatic tissue of the lung (3/6), lung (1/6), and stomach (1/6) of the infected mice; however, it was not detected in the liver (0/6) or kidney (0/6). In IN-inoculated mice, BVDV antigen was detected in the lung and mesenteric lymph nodes of one BVDV-infected mouse but was not detected in other tissues. The results of this study suggest that the spleen is the most reliable tissue for BVDV antigen detection using IHC in the IP-injected group. Our study demonstrates that mice can be infected by BVDV. This is the first report of BVDV infection in mice.
Sakamoto, M; Ishii, S; Nishioka, K; Shimada, K
In malnourished rats, nutritionally rehabilitated rats at various stages, and in well nourished rats, levels of serum complement after bacterial infection caused by Staphylococcus aureus, as well as tuberculin reactivity, were examined. The elevation of complement showed a peak 2--3 days after infection, herein called the first complement response. A reelevation occurred at a later stage, 7--14 days after infection, and is referred to as the second complement response. The first complement response was observed in all the rats after Staphylococcus aureus infection but it was greater in well nourished rats. In malnourished rats, only the first complement response was observed and the tuberculin reaction and second complement response were lacking. After 1 week of nutritional rehabilitation, 40% of the rats showed recovery of tuberculin responses and both the first and second complement responses were observed. Nutritionally rehabilitated rats treated longer than 2 weeks, together with the well nourished control rats, showed positive tuberculin reactivity. The second complement response was also observed in such rats when bacterial infection was severe but not with mild infection. PMID:511223
Santos, S V; Moura, J V L; Lescano, S A Z; Castro, J M; Ribeiro, M C S A; Chieffi, P P
Toxocara canis and Toxocara cati are nematode parasites in dogs and cats, respectively, transmitted by ingestion of embryonated eggs, transmammary and transplacental (T. canis) routes and paratenic host predation. Many parasites use mechanisms that change the behaviour of their hosts to ensure continued transmission. Several researchers have demonstrated behavioural changes in mouse models as paratenic hosts for T. canis. However, there have been no studies on behavioural changes in laboratory rats (Rattus norvegicus) experimentally infected with T. cati. This study investigated behavioural changes and muscle strength in male and female rats experimentally infected with T. cati or T. canis in acute and chronic phases of infection. Regardless of sex, rats infected with T. cati showed a greater decrease in muscle strength 42 days post infection compared to rats infected with T. canis. However, behavioural changes were only observed in female rats infected with T. canis.
Modrý, David; Hofmannová, Lada; Antalová, Zuzana; Sak, Bohumil; Kváč, Martin
The infectivity of Cryptosporidium muris and Cryptosporidium andersoni in various species of voles was studied using experimental infections. None of the experimental voles inoculated with 1 × 10(5) oocysts of Cryptosporidium spp. shed any oocysts during 40 DPI, except Brandt's vole (Lasiopodomys brandtii), which was susceptible to C. muris infection. Experiments confirmed the resistance of voles of the genus Microtus sensu stricto to infection with mammalian gastric cryptosporidia, which provides a new study model with prospects to more fully understand the processes involved in the phenomenon of host specificity of this group of protists.
Keating, M. Kelly; Spivey, Pamela; Lathrop, George W.; Powell, Nathaniel; Levin, Michael L.
Rickettsia slovaca is a tick-borne human pathogen that is associated with scalp eschars and neck lymphadenopathy known as tick-borne lymphadenopathy (TIBOLA) or Dermacentor-borne necrosis erythema and lymphadenopathy (DEBONEL). Originally, R. slovaca was described in Eastern Europe, but since recognition of its pathogenicity, human cases have been reported throughout Europe. European vertebrate reservoirs of R. slovaca remain unknown, but feral swine and domestic goats have been found infected or seropositive for this pathogen. Recently, a rickettsial pathogen identical to R. slovaca was identified in, and isolated from, the American dog tick, Dermacentor variabilis. In previous experimental studies, this organism was found infectious to guinea pigs and transovarially transmissible in ticks. In this study, domestic goats (Capra hircus) were experimentally inoculated with the North American isolate of this R. slovaca-like agent to assess their reservoir competence–the ability to acquire the pathogens and maintain transmission between infected and uninfected ticks. Goats were susceptible to infection as demonstrated by detection of the pathogen in skin biopsies and multiple internal tissues, but the only clinical sign of illness was transient fever noted in three out of four goats, and reactive lymphoid hyperplasia. On average, less than 5% of uninfected ticks acquired the pathogen while feeding upon infected goats. Although domestic goats are susceptible to the newly described North American isolate of R. slovaca, they are likely to play a minor role in the natural transmission cycle of this pathogen. Our results suggest that goats do not propagate the North American isolate of R. slovaca in peridomestic environments and clinical diagnosis of infection could be difficult due to the brevity and mildness of clinical signs. Further research is needed to elucidate the natural transmission cycle of R. slovaca both in Europe and North America, as well as to identify a
Carrion, Ricardo; Brasky, Kathleen; Mansfield, Keith; Johnson, Curtis; Gonzales, Monica; Ticer, Anysha; Lukashevich, Igor; Tardif, Suzette; Patterson, Jean
Lassa virus causes thousands of deaths annually in western Africa and is considered a potential biological weapon. In an attempt to develop a small nonhuman primate model of Lassa fever, common marmosets were subcutaneously inoculated with Lassa virus strain Josiah. This inoculation resulted in a systemic disease with clinical and morphological features mirroring those in fatal human Lassa infection: fever, weight loss, high viremia and viral RNA load in tissues, elevated liver enzymes, and severe morbidity between days 15 and 20. The most prominent histopathology findings included multifocal hepatic necrosis with mild inflammation and hepatocyte proliferation, lymphoid depletion, and interstitial nephritis. Cellular aggregates in regions of hepatocellular necrosis were largely composed of HAM56-positive macrophages, devoid of CD3-positive and CD20-positive cells, and characterized by marked reductions in the intensity of HLA-DP, DQ, DR staining. A marked reduction in the major histocompatibility complex class II expression was also observed in the lymph nodes. Immunophenotypic alterations in spleen included reductions in overall numbers of CD20-positive and CD3-positive cells and the disruption of lymphoid follicular architecture. These findings identify the common marmoset as an appropriate model of human Lassa fever and present the first experimental evidence that replication of Lassa virus in tissues is associated with alterations that would be expected to impair adaptive immunity.
Gutiérrez-Expósito, Daniel; Ortega-Mora, Luis M; Marco, Ignasi; Boadella, Mariana; Gortázar, Christian; San Miguel-Ayanz, José María; García-Lunar, Paula; Lavín, Santiago; Alvarez-García, Gema
Besnoitia besnoiti has been reported to affect cattle, wildebeest, kudu and impala, and B. tarandi other wild ruminants (caribou, reindeer, mule deer and musk ox), causing similar characteristic clinical signs and lesions. However, both Besnoitia species have been reported in different geographical areas and the link between the sylvatic and domestic life cycles of Besnoita spp. in wild ruminants and cattle remains unknown. The aim of this study was to evaluate the presence of specific antibodies against Besnoitia spp. in wild ruminants in Spain. A wide panel of sera from red deer (Cervus elaphus) (n=734), roe deer (Capreolus capreolus) (n=124), chamois (Rupicapra pyrenaica) (n=170) and mouflon (Ovis musimon) (n=20) collected from different locations of Spain was analyzed. Beef cattle were present in all sampled areas and, interestingly, bovine besnoitiosis has been widely reported in some of them (e.g., Pyrenees and Central Spain). Sera samples were first examined with an Enzyme-Linked Immunosorbent Assay (ELISA). For red deer and roe deer, the ELISA was standardized with positive and negative control sera from several Cervidae species (100% Se and 98% Sp). Chamois and mouflon sera samples were tested with a previously reported ELISA validated for bovine sera (97% Se and 95% Sp) using protein G as a conjugate. Positive results by ELISA were confirmed a posteriori with a tachyzoite-based Western blot. Sixty-one sera samples from red deer and 17 sera samples from roe-deer were seropositive or doubtful by ELISA. All samples from mouflon were seronegative and 15 sera samples from chamois were considered doubtful. B. besnoiti exposure was only confirmed clearly by Western blot in one red deer and one roe deer from the Spanish Pyrenees where the disease is traditionally endemic. This is the first serological report of Besnoitia spp. infection carried out in European wild ruminants and the results show that specific antibodies are present at least in red deer and roe
Corner, L A L; Costello, E; Lesellier, S; O'Meara, D; Sleeman, D P; Gormley, E
The aim was to develop an endobronchial infection procedure for the study of Mycobacterium bovis infection in badgers. The badgers were anaesthetised and a cannula was passed per os to the tracheal bifurcation. When in place 1 ml of M. bovis suspension was inoculated. Three concentrations of M. bovis suspension were used; <10 colony forming units (cfu), approximately 10(2) cfu and approximately 3 x 10(3) cfu. The badgers were examined at three weekly intervals for clinical signs of disease and a tracheal aspirate was collected at each examination. The badgers were euthanased 17 weeks post infection (pi) and at the post mortem examination a wide range of tissues were examined for gross and histopathological lesions of tuberculosis and cultured for M. bovis. A sample of bronchial alveolar lavage (BAL) fluid was collected at post mortem for culture. At post mortem examination 17 weeks after infection, gross and histopathological lesions of tuberculosis were observed in all badgers inoculated with the high and medium dose and 1/3 inoculated with the low dose. M. bovis was recovered from all inoculated badgers. Infection in the high dose group was more widely disseminated than in the other groups. The number of sites with gross and histopathological lesions increased with increasing dose of M. bovis. All tracheal aspirates were negative on culture and only one BAL, collected from a badger of the high dose group, was positive on culture. No clinical signs due to the experimental infection were observed. The endobronchial route of inoculation is an effective route for establishing experimental infection, and could be used for studies of tuberculosis pathogenesis, immunology of M. bovis infection in badgers and for challenging badgers in vaccine protection studies. Badgers appeared to be very susceptible to infection by this procedure even with a dose of < 10 cfu but appear to control and limit the resulting infection.
Dubey, J P; Lindsay, D S; Saville, W J A
Sarcocystis neurona is the most important cause of a neurologic disease of horses, equine protozoal myeloencephalitis (EPM). Cats and other carnivores can act as its intermediate hosts and horses are aberrant hosts. Little is known of the sero-epidemiology of S. neurona infections in cats. In the present study, antibodies to S. neurona were evaluated by the S. neurona agglutination test (SAT). Cats fed sporocysts from the feces of naturally infected opossums or inoculated intramuscularly with S. neurona merozoites developed high levels (> or =1:4000) of SAT antibodies. Antibodies to S. neurona were not found in a cat inoculated with merozoites of the closely related parasite, Sarcocystis falcatula. These results should be useful in studying sero-epidemiology of S. neurona infections in cats.
Chen, S C; You, S H; Liu, C Y; Chio, C P; Liao, C M
The aim of this work was to use experimental infection data of human influenza to assess a simple viral dynamics model in epithelial cells and better understand the underlying complex factors governing the infection process. The developed study model expands on previous reports of a target cell-limited model with delayed virus production. Data from 10 published experimental infection studies of human influenza was used to validate the model. Our results elucidate, mechanistically, the associations between epithelial cells, human immune responses, and viral titres and were supported by the experimental infection data. We report that the maximum total number of free virions following infection is 10(3)-fold higher than the initial introduced titre. Our results indicated that the infection rates of unprotected epithelial cells probably play an important role in affecting viral dynamics. By simulating an advanced model of viral dynamics and applying it to experimental infection data of human influenza, we obtained important estimates of the infection rate. This work provides epidemiologically meaningful results, meriting further efforts to understand the causes and consequences of influenza A infection.
Attempts were made to develop an animal model for Cyclospora cayetanensis to identify a practical laboratory host for studying human cyclosporiasis. Oocysts collected from stool of infected humans in the United States, Haiti, Guatemala, Peru and Nepal were held in potassium dich...
Attemps were made to develop an animal model for Cyclospora cayetanensis to identify a practical laboratory host for studing human cyclosporiasis. Oocysts collected from stool of infected humans in the United States, Haiti, Guatemala, Peru, and Nepal were held in potassium dichro...
Malizos, K; Sarma, J; Seaton, R A; Militz, M; Menichetti, F; Riccio, G; Gaudias, J; Trostmann, U; Pathan, R; Hamed, K
Osteomyelitis is a serious infection predominantly caused by Gram-positive bacteria, including methicillin-resistant Staphylococcus aureus (MRSA). Orthopaedic device-related infections are complex and require a careful combination of surgical intervention and antimicrobial therapy. Daptomycin, a cyclic lipopeptide, effectively penetrates soft tissue and bone and demonstrates rapid concentration-dependent bactericidal activity against Gram-positive pathogens. This retrospective, non-interventional study evaluated clinical outcomes in patients with osteomyelitis or orthopaedic device infections treated with daptomycin from the European Cubicin® Outcomes Registry and Experience (EU-CORE(SM)) study. Patients were treated between January 2006 and April 2012, with follow-up to 2014. Clinical outcomes were assessed as success (cured or improved), failure or non-evaluable. Of 6,075 patients enrolled, 638 (median age, 63.5 years) had primary infections of osteomyelitis or orthopaedic device infections, 224 had non-prosthetic osteomyelitis, 208 had osteomyelitis related to a permanent or temporary prosthetic device, and 206 had orthopaedic device infections. The most commonly isolated pathogen was S. aureus (214 [49.1 %]; 24.8 % were MRSA). Overall, 455 (71.3 %) patients had received previous antibiotic therapy. Patients underwent surgical interventions, including tissue (225 [35.3 %]) and bone (196 [30.7 %]) debridement, as part of their treatment. Clinical success rates were 82.7 % and 81.7 % in S. aureus and coagulase-negative staphylococcal infections. Adverse events (AEs) and serious AEs assessed as possibly related to daptomycin were observed in 6.7 % and 1.9 % of patients, respectively. Daptomycin was discontinued by 5.5 % of patients due to AEs and 10 (1.6 %) deaths were reported. In conclusion, daptomycin was effective and safe in patients with osteomyelitis or orthopaedic device infections.
Boothby, J T; Schore, C E; Jasper, D E; Osburn, B I; Thomas, C B
This study characterized the immune responses in four vaccinated and four control cows in response to vaccination and experimental intramammary inoculation with Mycoplasma bovis. Specific antibody responses occurred in serum and milk in response to vaccination and experimental infection. Lymphocytes from peripheral blood, but not from the mammary gland of vaccinated cows had increased responsiveness to mitogens. No lymphocytes tested were responsive to M. bovis antigen. Both vaccination and experimental infection resulted in skin test reactivity. These results imply that vaccination results in immune responses which may alter the course of experimental M. bovis mastitis, but may contribute to cellular inflammation. PMID:3167718
Lesellier, Sandrine; Corner, Leigh; Costello, Eamon; Sleeman, Paddy; Lyashchenko, Konstantin; Greenwald, Rena; Esfandiari, Javan; Singh, Mahavir; Hewinson, R Glyn; Chambers, Mark; Gormley, Eamonn
European badgers (Meles meles) are considered to be an important reservoir of infection for Mycobacterium bovis and are implicated in the transmission of tuberculosis to cattle in Ireland and Great Britain. Accurate tests are required for tuberculosis surveillance in badger populations and to provide a basis for the development of strategies, including vaccination, to reduce the incidence of the infection. In this study, we have developed an endobronchial M. bovis infection model in badgers in which we measured cell-mediated immune and serological responses for up to 24 weeks post-infection. Groups of badgers were subjected to necropsy at 6-week intervals and the gross lesion severity status compared with immune responses measured in blood samples taken throughout the course of the study. The panel of antigens included bovine and avian tuberculins (PPD) as well as single antigens, ESAT-6, CFP-10, MPB70, Rv3019c, Rv3873, Rv3878 and Rv3879, all known to be recognised by the immune system in other animal models of tuberculosis infection. Our results demonstrated that M. bovis infected badgers responded to specific antigens as early as 6 weeks post-infection, consistent with the presence of visible lesions. The data also revealed unique patterns of antigen recognition with high levels of PBMC proliferation in the presence of CFP-10 but low proliferation levels with ESAT-6. Using a multi-antigen print immunoassay (MAPIA), we were able to confirm that MPB83 is the dominant antigen recognised by serum antibodies in infected badgers.
Infection models are useful for studying host responses to infection to aid in the development of diagnostic tools and vaccines. The majority of experimental models for ruminants have utilized an oral inoculation of live Mycobacterium avium subsp. paratuberculosis (MAP) in order to establish infecti...
Animal models are useful for studying host responses to infection and aid in the development of diagnostic tools and vaccines. The current study was designed to compare the effects of different methods of experimental infection: Oral (Mycobacterium avium subsp. parauberculosis (MAP) strain K-10; Or...
Animal models are useful for studying host responses to infection and aid in the development of diagnostic tools and vaccines. The current study was designed to compare the effects of different methods of experimental infection: Oral (Mycobacterium avium subsp. parauberculosis (MAP) strain K-10; Or...
Neospora caninum is a major cause of abortion in cattle, but the reasons why some animals abort and not others remain unclear. Most of the N. caninum experimental primary infections in cattle late in gestation, after 120 days of pregnancy, results in birth of full-term congenitally infected fetuses....
Costa, Dirceu Joaquim; Carvalho, Rayssa M. de Araujo; Abbehusen, Melissa; Teixeira, Clarissa; Pitombo, Maiana; Trigo, Joelma; Nascimento, Flávia; Amorim, Lucilene; Abreu-Silva, Ana Lucia; do Socorro Pires Cruz, Maria; Miranda, José Carlos; Fukutani, Kyoshi; de Oliveira, Camila I.; Barral, Aldina; Barral-Netto, Manoel; Brodskyn, Cláudia
Background Canine Visceral Leishmaniasis (CVL) is a zoonotic disease caused by Leishmania infantum, transmitted by the bite of Lutzomyia longipalpis sand flies. Dogs are the main domestic reservoir of the parasite. The establishment of an experimental model that partially reproduces natural infection in dogs is very important to test vaccine candidates, mainly regarding those that use salivary proteins from the vector and new therapeutical approaches. Methodology/Principal Findings In this report, we describe an experimental infection in dogs, using intradermal injection of Leishmania infantum plus salivary gland homogenate (SGH) of Lutzomyia longipalpis. Thirty-five dogs were infected with 1×107 parasites combined with five pairs of Lutzomyia longipalpis salivary glands and followed for 450 days after infection and clinical, immunological and parasitological parameters were evaluated. Two hundred and ten days after infection we observed that 31,4% of dogs did not display detectable levels of anti-Leishmania antibodies but all presented different numbers of parasites in the lymph nodes. Animals with a positive xenodiagnosis had at least 3,35×105 parasites in their lymph nodes. An increase of IFN-γ and IL-10 levels was detected during infection. Twenty two percent of dogs developed symptoms of CVL during infection. Conclusion The infection model described here shows some degree of similarity when compared with naturally infected dogs opening new perspectives for the study of CVL using an experimental model that employs the combination of parasites and sand fly saliva both present during natural transmission. PMID:23577121
Vannucci, Fabio Augusto; Borges, Elizabeth Lage; de Oliveira, Juliana Saes Vilaça; Guedes, Roberto Mauricio Carvalho
The objective of this study was to evaluate the intestinal absorption and histomorphometry of hamsters experimentally infected with Lawsonia intracellularis and correlate these parameters with severity of infection based on immunohistochemistry. Sixty hamsters were equally divided into control and inoculated groups which were orally infected with intestinal mucosa homogenate from pigs naturally infected with L. intracellularis. The intestinal absorption of glucose, sodium, potassium and chloride was evaluated in live animals (25 inoculated and 25 control) on day 26 after inoculation. In this procedure, a standard solution was infused into the cranial jejunum and collected at the terminal ileum. The experimental infection was confirmed by gross and histopathological examination and L. intracellularis antigen labeling by immunohistochemistry. Histomorphometry analysis demonstrated positive correlation between intestinal crypt depth and severity of infection based on immunohistochemistry. Infected animals had significantly lower intestinal absorption of glucose, potassium and chloride. These results indicate a lower intestinal absorption as an important mechanism of diarrhea in hamsters experimentally infected with L. intracellularis. Therefore, malabsorption should be considered as the main mechanism involved in the physiopathology of the diarrhea in L. intracellularis infected animals.
Petranyi, G; Meingassner, J G; Mieth, H
The allylamine derivative terbinafine is the first antifungal agent with primary fungicidal properties against dermatophytes which acts systemically after oral application as well as locally after topical application. Comparative oral studies carried out with griseofulvin and ketoconazole in model infections such as guinea pig trichophytosis and microsporosis revealed terbinafine to be superior to the reference compounds both clinically and mycologically. An excellent antimycotic activity of terbinafine was also demonstrable after topical treatment of guinea pig dermatophytoses caused by Trichophyton mentagrophytes or Microsporum canis. Results of comparative chemotherapeutic studies carried out with econazole and tolnaftate demonstrated superior efficacy of terbinafine in the treatment of both trichophytosis and microsporosis. Skin infections of guinea pigs caused by Candida albicans and vaginal candidiasis in rats proved to be responsive to a topical application of terbinafine also. However, the reference compounds, clotrimazole and miconazole, exhibited activity superior to that of terbinafine in both models. PMID:3435103
Resende, Deisy V; Lages-Silva, Eliane; Assis, Dnieber C; Prata, Aluízio; Oliveira-Silva, Márcia B
Extraintestinal cystoisosporosis by Cystoisospora belli has already been reported in HIV/AIDS patients, generally involving preferential invasion of mesenteric and trachaeobronchial lymph nodes, liver and spleen by unizoic cysts of this parasite, which may infect macrophages. To test this hypothesis, murine and human macrophages were exposed to sporozoites of C. belli and cultures were observed daily after contact with these cells. The parasites penetrated and multiplied by endodyogeny in both cell types and inserted themselves inside perinuclear vacuoles. After 48 h, extracellular parasites were removed from macrophage cultures and incubated in Monkey Kidney Rhesus cells (MK2) where there was intense multiplication. This is the first report of infection of macrophages by this parasite, which supports the hypothesis that these could act as C. belli host cells in extraintestinal sites.
El-Nouby, Kholoud A; Hamouda, Hala E; Abd El Azeem, Mona A; El-Ebiary, Ahmad A
The effect of sodium benzoate (SB) on the pathogenesis of Hymenolepis nana (H. nana) and its neurological manifestations was studied in the present work. One hundred and thirty five mice were classified into three groups. GI: received SB alone. GII: received SB before & after infection with H. nana and GIII: infected with H. nana. All groups were subjected to parasitological, histopathological, immunohistochemical and biochemical assays. The results revealed a significant decrease in IL-4 serum level with a significant increase in gamma amino butyric acid (GABA) and decrease in zinc brain levels in GI, while GII showed non significant increase in IL-4 level that resulted in a highly significant increase in the mean number of cysticercoids and adult worms with delayed expulsion as compared to GIII. This was reflected on histopathological and immunohistochemical changes in the brain. Also, there was a highly significant increase in GABA and decrease in zinc brain levels in GII to the degree that induced behavioral changes. This emphasizes the possible synergistic effect of SB on the neurological manifestations of H. nana and could, in part, explain the increased incidence of behavioral changes in children exposed to high doses of SB and unfortunately have H. nana infection.
Carrasco, N; Arzul, I; Chollet, B; Robert, M; Joly, J P; Furones, M D; Berthe, F C J
Paracartia grani (Copepoda) has been identified as a potential intermediate host in the life cycle of Marteilia refringens, a paramyxean parasite infecting flat oysters. However, no intermediate host has yet been identified for Marteilia maurini that infects mussels. A better understanding of the life cycle of these two Marteilia types would clarify their taxonomic relationship and hypothesized co-specificity. For this purpose, experimental infections of copepods, P. grani, were performed using naturally infected flat oysters and mussels. Infection patterns were different depending whether copepods were infected from oysters or mussels. M. maurini did not proliferate in copepods while M. refringens rapidly proliferated in infected copepods. Previously unrecognized developmental stages of M. refringens were found during this study.
Rondelaud, D; Teukeng, F F Djuikwo; Vignoles, P; Dreyfuss, G
Experimental infections of Lymnaea glabra (two populations) with Fasciola hepatica were carried out during seven successive snail generations, to determine if prevalence and intensity of snail infection increased over time through descendants of snails already infected with F. hepatica. Controls were descendants coming from uninfected parents and infected according to the same protocol. No larval forms were found in the bodies of control snails coming from uninfected parents. In contrast, prevalence and intensity of F. hepatica infection in snails originating from infected parents progressively increased from the F2 or F3 to the F6 generation of L. glabra. In another experiment carried out with the F7 generations of L. glabra and a single generation of Galba truncatula (as controls), the prevalence of F. hepatica infection and the total number of cercariae were lower in L. glabra (without significant differences between both populations). If the number of cercariae shed by infected snails was compared to overall cercarial production noted in snails containing cercariae but dying without emission, the percentage was greater in G. truncatula (69% instead of 52-54% in L. glabra). Even if most characteristics of F. hepatica infection were lower in L. glabra, prevalence and intensity of parasite infection increased with snail generation when tested snails came from infected parents. This mode of snail infection with F. hepatica suggests an explanation for cases of fasciolosis occurring in cattle-breeding farms where paramphistomosis is lacking and G. truncatula is absent.
A european class is, in France, a class in which we teach a subject in a foreign language, for example science in English. I led, in my European class, during a seven weeks session, group work activities about different participating science actions. There were groups composed of three or four 16 years old students. Each group chose one type of participating science activity among : - Leading a visioconference with an IODP mission on board the Joides Resolution. - Being part of a "science songs community" with Tom Mc Fadden They divided the work and some of them studied the websites and contacted the actors to present the pedagogical or scientific background of their subject. Others had a concrete production like the organization of a visioconference with the Joides Resolution or the creation of a pedagogical song about geology. I will present some results of their work and explain the students motivation linked to this active learning method.
Nair, Arathy D S; Cheng, Chuanmin; Ganta, Chanran K; Sanderson, Michael W; Alleman, Arthur R; Munderloh, Ulrike G; Ganta, Roman R
Dogs acquire infections with the Anaplasmataceae family pathogens, E. canis, E. chaffeensis, E. ewingii, A. platys and A. phagocytophilum mostly during summer months when ticks are actively feeding on animals. These pathogens are also identified as causing diseases in people. Despite the long history of tick-borne diseases in dogs, much remains to be defined pertaining to the clinical and pathological outcomes of infections with these pathogens. In the current study, we performed experimental infections in dogs with E. canis, E. chaffeensis, A. platys and A. phagocytophilum. Animals were monitored for 42 days to evaluate infection-specific clinical, hematological and pathological differences. All four pathogens caused systemic persistent infections detectible throughout the 6 weeks of infection assessment. Fever was frequently detected in animals infected with E. canis, E. chaffeensis, and A. platys, but not in dogs infected with A. phagocytophilum. Hematological differences were evident in all four infected groups, although significant overlap existed between the groups. A marked reduction in packed cell volume that correlated with reduced erythrocytes and hemoglobin was observed only in E. canis infected animals. A decline in platelet numbers was common with E. canis, A. platys and A. phagocytophilum infections. Histopathological lesions in lung, liver and spleen were observed in all four groups of infected dogs; infection with E. canis had the highest pathological scores, followed by E. chaffeensis, then A. platys and A. phagocytophilum. All four pathogens induced IgG responses starting on day 7 post infection, which was predominantly comprised of IgG2 subclass antibodies. This is the first detailed investigation comparing the infection progression and host responses in dogs after inoculation with four pathogens belonging to the Anaplasmataceae family. The study revealed a significant overlap in clinical, hematological and pathological changes resulting from the
Edsall, Geoffrey; Gaines, Sidney; Landy, Maurice; Tigertt, W. D.; Sprinz, Helmuth; Trapani, R.-J.; Mandel, Adrian D.; Benenson, A. S.
A disease resembling human typhoid fever has been induced by feeding live cultures of Salmonella typhosa to young chimpanzees, thus confirming the classical reports of Grünbaum and of Metchnikoff and Besredka. Detailed clinical observations, results of stool and blood cultures, and serological studies have confirmed the impression that the disease produced in chimpanzees closely resembles the mild form of human typhoid fever frequently seen in childhood. Gross and histologic examination of intestines, mesenteric lymph nodes, liver, spleen, and other organs of orally infected chimpanzees has demonstrated that the pathological findings are essentially indistinguishable from those seen in mild typhoid fever in man. The clinical spectrum of disease seen in chimpanzees ranged from moderately severe illness, through transitory illness, to afebrile infection with or without bacteriemia (but invariably with an antibody response), occasionally leading to the development of persisting biliary infection and the carrier state. Thus the range of illness observed in chimpanzees resembled that seen in man, except that the severe and complicated forms of typhoid fever were not observed in the chimpanzee. A reason for this difference is proposed and discussed. In contrast to the limitations imposed upon the interpretation of human epidemiologic observations, it has been possible to demonstrate in the chimpanzee that clinical variation in disease pattern from animal to animal may occur despite the administration of the same dose of the same bacterial strain simultaneously to an entire group of animals under study; in other words, variation in clinical pattern is dependent on inherent, non-specific host factors as well as on dose, strain or preceding state of immunity. Variation in dose and in challenge strain of S. typhosa employed also appeared to have an effect upon the likelihood of producing febrile as against afebrile infection in chimpanzees. The dose required to produce
Walton, Thomas E.; Johnson, Karl M.
Two groups of four dairy cows (Bos taurus) were infected subcutaneously with the epizootic Venezuelan equine encephalomyelitis virus (VEE) strains MF-8 and San Pelayo, respectively. Animals experienced no clinical illness, but all developed significant neutropenia. Virus was recovered once each from the blood of three animals but did not exceed 102.2 SMICLD50 (Suckling mouse intracerebral lethal dose50)/ml. Specific neutralizing antibodies appeared in the serum of all animals, but there were no significant differences in titers against different naturally occurring VEE subtypes. Dairy cattle thus appear to play no role in virus transmission during VEE epizootics but may serve as retrospective immunological sentinels of virus activity. PMID:4564396
Lim, Sun; Park, Sang-Ik; Ahn, Kyu-Sung; Oh, Dae-Sung; Shin, Sung-Shik
To evaluate the efficacy of ronidazole for treatment of Tritrichomonas foetus infection, 6 Tritrichomonas-free kittens were experimentally infected with a Korean isolate of T. foetus. The experimental infection was confirmed by direct microscopy, culture, and single-tube nested PCR, and all cats demonstrated trophozoites of T. foetus by day 20 post-infection in the feces. From day 30 after the experimentally induced infection, 3 cats were treated with ronidazole (50 mg/kg twice a day for 14 days) and 3 other cats received placebo. Feces from each cat were tested for the presence of T. foetus by direct smear and culture of rectal swab samples using modified Diamond's medium once a week for 4 weeks. To confirm the culture results, the presence of T. foetus rRNA gene was determined by single-tube nested PCR assay. All 3 cats in the treatment group receiving ronidazole showed negative results for T. foetus infection during 2 weeks of treatment and 4 weeks follow-up by all detection methods used in this study. In contrast, rectal swab samples from cats in the control group were positive for T. foetus continuously throughout the study. The present study indicates that ronidazole is also effective to treat cats infected experimentally with a Korean isolate of T. foetus at a dose of 50 mg/kg twice a day for 14 days.
Doha, Said A; Shehata, Magdi G; Fahmy, Adel R; Samy, Abdallah M
Cutaneous leishmaniasis (CL) is a neglected clinical form that is quite prevalent in Eastern North parts of the country in Sinai Peninsula. Leishmania tropica was identified by previous reports as the causative agent responsible for viscerotropic infections in-patients and experimental animals. Here, we reported the viscerotropic infections from naturally infected rodent Gerbillus pyramidum floweri collected from North-Sinai. Footpad and tail lesions, spleenomegaly, and malformed dark-colored spleen were the characteristic CL symptoms. The spleen of the rodent found positive to amastigote impression smear. ITS-1 DNA was sequenced and revealed 100% identity of the strain in the current study to the other L. tropica sequences identified from the patients with the suspected CL and inhabited the same study area. The current findings confirmed the susceptibility of gerbil to L. tropica, and raise the concerns for the role of rodents as accidental host suffering the infections. The susceptibility of wild and experimental rodents to the same L. tropica strain was also investigated; BALB/c and G. pyramidum were more susceptible to L. tropica (24.33 ± 4.37 and 25 ± 4.58 days post-infection, respectively). Similar viscerotropic pathologies were reported in experimental infection of only golden hamster (≈ 120 days post-infection), and G. p. floweri (≈ 160 days post-infection).
Corba, J; Spaldonová, R
Results are presented on the effect of immunosuppressive substances such as chlorambucil, cyclophosphamide, azathioprine, amethopterine and a cortizone derivate of betamethasone, on the development of Fasciola hepatica in the rat. The suppression of the immune response of the host to immunosuppressants was reflected in an earlier start of migration of the flukes to the common bile duct, and in an earlier onset of egg production as compared with that in the controls. Of the substances employed, cyclophosphamide and betamethasone were the most effective ones within the period from week 2--6 p.i., which is the time during which the migration of the flukes in the liver parenchyma is highest. Pathological changes in the liver of the animals were less marked than those of the infected controls. Evidence was obtained on an increased pathogenicity of infective larval flukes causing a higher mortality of the hosts in comparison with that of the control animals. On the other hand, the administration of immunosuppressants did neither influence the total number of developed flukes nor the appearance of eosinophilia in the peripheral blood of the treated animals.
Gajadhar, A A; Measures, L; Forbes, L B; Kapel, C; Dubey, J P
Laboratory-reared animals were used to assess the susceptibility of seals (Halichoerus grypus) to Toxoplasma gondii infection. Four seals were each orally inoculated with 100 or 10,000 oocysts of T. gondii (VEG strain), and another 4 seals served as negative controls. Occasionally, mild behavioral changes were observed in all inoculated seals but not in control animals. A modified agglutination test revealed the presence of antibodies to T. gondii in sera collected from inoculated seals and mice inoculated as controls. No evidence of the parasite was found on an extensive histological examination of seal tissues, and immunohistochemical staining of tissue sections from inoculated seals revealed a single tissue cyst in only 1 seal. Control mice inoculated with 10 oocysts from the same inoculum given to seals became serologically and histologically positive for T. gondii. Cats that were fed brain or muscle tissue collected from inoculated seals passed T. gondii oocysts in feces. This study demonstrates that T. gondii oocysts can establish viable infection in seals and supports the hypothesis that toxoplasmosis in marine mammals can be acquired from oocysts in surface water runoff and sewer discharge.
Pérez del Villar, Luis; Vicente, Belén; Galindo-Villardón, Purificación; Castellanos, Andrés; Pérez-Losada, Jesús; Muro, Antonio
Most Schistosoma mansoni experimental infections are developed in several inbred strains of Mus musculus as definitive host. In contrast, Mus spretus is unexplored in Schistosoma infection studies. Mus spretus provides a high variation of immunological phenotypes being an invaluable tool for genetic studies and gene mapping. The aim of this study is to characterize hematological and immunological responses against Schistosoma mansoni infection in Mus spretus (SPRET/EiJ strain) vs. Mus musculus (CD1 strain) mice. Nine weeks after cercarial exposure, animals were perfused and the parasite burden was assessed. The parasitological data suggests that SPRET/EiJ mice tolerate higher parasite loads compared to CD1 strain. In addition, hematological parameters measured in Mus spretus group showed a significant increase in granulocytes population in early stages of infection compared to the CD1 cohort. Meanwhile, CD1 presented higher levels of lymphocytes and IgG1 in the late stages of S. mansoni experimental infection. PMID:23985166
Valero, Yulema; García-Alcázar, Alicia; Esteban, M Ángeles; Cuesta, Alberto; Chaves-Pozo, Elena
Antimicrobial peptides (AMPs) have a crucial role in the fish innate immune response, being considered a fundamental component of the first line of defence against pathogens. Moreover, AMPs have not been studied in the fish gonad since this is used by some pathogens as a vehicle or a reservoir to be transmitted to the progeny, as occurs with nodavirus (VNNV), which shows vertical transmission through the gonad and/or gonadal fluids, but no study has looked into the gonad of infected fish. In this framework, we have characterized the antimicrobial response triggered by VNNV in the testis of European sea bass, a very susceptible species of the virus, and in the gilthead seabream, which acts as a reservoir, both in vivo and in vitro, and compared with that present in the serum and brain (target tissue of VNNV). First, our data show a great antiviral response in the brain of gilthead seabream and in the gonad of European sea bass. In addition, for the first time, our results demonstrate that the antimicrobial activities (complement, lysozyme and bactericidal) and the expression of AMP genes such as complement factor 3 (c3), lysozyme (lyz), hepcidin (hamp), dicentracin (dic), piscidin (pis) or β-defensin (bdef) in the gonad of both species are very different, but generally activated in the European sea bass, probably related with the differences of susceptibility upon VNNV infection, and even differs to the brain response. Furthermore, the in vitro data suggest that some AMPs are locally regulated playing a local immune response in the gonad, while others are more dependent of the systemic immune system. Data are discussed in the light to ascertain their potential role in viral clearance by the gonad to avoid vertical transmission.
McLean, R G; Crans, W J; Caccamise, D F; McNelly, J; Kirk, L J; Mitchell, C J; Calisher, C H
To study the susceptibility of wading birds to eastern equine encephalitis (EEE) virus and to determine their potential as reservoir or amplifying hosts, fledgling glossy ibises (Plegadis falcinellus) and snowy egrets (Egretta thula) were captured in New Jersey (USA) and shipped to Colorado (USA) where they were experimentally inoculated with EEE virus. All 16 snowy egrets and 14 (93%) of 15 of the glossy ibises inoculated became viremic with moderate titers, and all survivors developed neutralizing antibody. Six ibises and two egrets died during the first week after inoculation, and EEE virus was isolated from the tissues of three birds. Our experimental results support field evidence about the relative involvement of glossy ibises and snowy egrets in the epizootiology of EEE virus in New Jersey.
Carlson, CM; Schneider, Jay R.; Pedersen, Janice C.; Heisey, Dennis M.; Johnson, Christopher J.
Meadow voles (Microtus pennsylvanicus) are permissive to chronic wasting disease (CWD) infection, but their susceptibility to other transmissible spongiform encephalopathies (TSEs) is poorly characterized. In this initial study, we intracerebrally challenged 6 meadow voles with 2 isolates of sheep scrapie. Three meadow voles acquired a TSE after the scrapie challenge and an extended incubation period. The glycoform profile of proteinase K-resistant prion protein (PrP(res)) in scrapie-sick voles remained similar to the sheep inocula, but differed from that of voles clinically affected by CWD. Vacuolization patterns and disease-associated prion protein (PrP(Sc)) deposition were generally similar in all scrapie-affected voles, except in the hippocampus, where PrP(Sc) staining varied markedly among the animals. Our results demonstrate that meadow voles can acquire a TSE after intracerebral scrapie challenge and that this species could therefore prove useful for characterizing scrapie isolates.
Beaman, Blaine L.
Several strains of Nocardia that varied from virulent to avirulent were injected intraperitoneally into young mice. Histological and ultrastructural analysis of the resultant infections revealed that the bacteria and the lesions they induced were different depending upon the strain of organism used. Further, the morphological and tinctorial characteristics of the bacteria grown in vitro changes during growth in vivo. These observations strongly suggested that chemical and physical alterations occurred in the cell envelope of the Nocardia when grown in mice. Electron microscopy confirmed that significant structural modification occurred, especially in the cell envelope, when the nocardial cells established themselves within the host tissue. It was shown that the least virulent strain exhibited the most dramatic changes whereas the most virulent organism appeared to be affected the least. Images PMID:4584055
Carlson, Christina M.; Schneider, Jay R.; Pedersen, Joel A.; Heisey, Dennis M.; Johnson, Christopher J.
Meadow voles (Microtus pennsylvanicus) are permissive to chronic wasting disease (CWD) infection, but their susceptibility to other transmissible spongiform encephalopathies (TSEs) is poorly characterized. In this initial study, we intracerebrally challenged 6 meadow voles with 2 isolates of sheep scrapie. Three meadow voles acquired a TSE after the scrapie challenge and an extended incubation period. The glycoform profile of proteinase K-resistant prion protein (PrPres) in scrapie-sick voles remained similar to the sheep inocula, but differed from that of voles clinically affected by CWD. Vacuolization patterns and disease-associated prion protein (PrPSc) deposition were generally similar in all scrapie-affected voles, except in the hippocampus, where PrPSc staining varied markedly among the animals. Our results demonstrate that meadow voles can acquire a TSE after intracerebral scrapie challenge and that this species could therefore prove useful for characterizing scrapie isolates. PMID:25673912
Carlson, Christina M; Schneider, Jay R; Pedersen, Joel A; Heisey, Dennis M; Johnson, Christopher J
Meadow voles (Microtus pennsylvanicus) are permissive to chronic wasting disease (CWD) infection, but their susceptibility to other transmissible spongiform encephalopathies (TSEs) is poorly characterized. In this initial study, we intracerebrally challenged 6 meadow voles with 2 isolates of sheep scrapie. Three meadow voles acquired a TSE after the scrapie challenge and an extended incubation period. The glycoform profile of proteinase K-resistant prion protein (PrP(res)) in scrapie-sick voles remained similar to the sheep inocula, but differed from that of voles clinically affected by CWD. Vacuolization patterns and disease-associated prion protein (PrP(Sc)) deposition were generally similar in all scrapie-affected voles, except in the hippocampus, where PrP(Sc) staining varied markedly among the animals. Our results demonstrate that meadow voles can acquire a TSE after intracerebral scrapie challenge and that this species could therefore prove useful for characterizing scrapie isolates.
Rivers, T M; Berry, G P
1. Rabbits and guinea pigs are susceptible to psittacosis virus introduced intracerebrally. By means of brain to brain passages in these animals the active agent is capable of propagation indefinitely. 2. Serial passages of the virus through rabbits and guinea pigs do not cause the active agent to lose its pathogenicity for parrots and mice. 3. The chief clinical evidences of infection in rabbits and guinea pigs following intracranial inoculation of the virus are fever and loss of weight. The pathological changes are characterized by a mild meningo-encephalitis, and fatty degeneration, focal necrosis, and infarction of the liver. 4. Rabbits upon recovery from an attack of psittacosis are actively immune. 5. Two strains of virus, human and parrot, were found to be immunologically similar. 6. No evidence was obtained to show that human convalescent serum possesses an appreciable amount of neutralizing substances.
McClain, D J; Summers, P L; Pratt, W D; Davis, K J; Jennings, G B
Due to the lack of an animal model, previous studies of sandfly fever have relied upon human challenge trials. We examined the infectivity and potential pathogenicity of sandfly fever virus in cynomolgus monkeys (Macaca fascicularis). Three different preparations of sandfly fever virus. Sicilian strain, and a placebo were compared by different routes of administration. The most notable postchallenge clinical event was a decrease in lymphocytes in the intramuscularly challenged monkeys. Plaque-reduction neutralization responses peaked earlier in animals challenged intravenously as compared with those in animals challenged intramuscularly. There was no evidence for neurotropism or meningeal inflammation. Sandfly fever virus was infectious for cynomolgus monkeys, but produced no detectable clinical disease that might serve as a marker for animal modeling studies. On the other hand, the preclinical data provide supportive evidence for safe parenteral administration of a Sicilian strain of sandfly fever virus inoculum to humans as a challenge model for sandfly fever disease.
Lacroux, Caroline; Perrin-Chauvineau, Cécile; Corbière, Fabien; Aron, Naima; Aguilar-Calvo, Patricia; Torres, Juan Maria; Costes, Pierrette; Brémaud, Isabelle; Lugan, Séverine; Schelcher, François; Barillet, Francis; Andréoletti, Olivier
In goats, several field studies have identified coding mutations of the gene encoding the prion protein (I/M142, N/D146, S/D146, R/Q211, and Q/K222) that are associated with a lower risk of developing classical scrapie. However, the data related to the levels of resistance to transmissible spongiform encephalopathies (TSEs) of these different PRNP gene mutations are still considered insufficient for developing large-scale genetic selection against scrapie in this species. In this study, we inoculated wild-type (WT) PRNP (I142R154R211Q222) goats and homozygous and/or heterozygous I/M142, R/H154, R/Q211, and Q/K222 goats with a goat natural scrapie isolate by either the oral or the intracerebral (i.c.) route. Our results indicate that the I/M142 PRNP polymorphism does not provide substantial resistance to scrapie infection following intracerebral or oral inoculation. They also demonstrate that H154, Q211, and K222 PRNP allele carriers are all resistant to scrapie infection following oral exposure. However, in comparison to WT animals, the H154 and Q211 allele carriers displayed only moderate increases in the incubation period following i.c. challenge. After i.c. challenge, heterozygous K222 and a small proportion of homozygous K222 goats also developed the disease, but with incubation periods that were 4 to 5 times longer than those in WT animals. These results support the contention that the K222 goat prion protein variant provides a strong but not absolutely protective effect against classical scrapie.
Sakai, Renata K; Costa, Francisco B; Ueno, Tatiana E H; Ramirez, Diego G; Soares, João F; Fonseca, Adivaldo H; Labruna, Marcelo B; Barros-Battesti, Darci M
Amblyomma dubitatum engorged females, naturally infected by Rickettsia bellii, were used to establish a laboratory colony. Larvae, nymphs, and adults were exposed to two strains of Rickettsia rickettsii by feeding on needle-inoculated guinea pigs, and thereafter reared on uninfected guinea pigs. After acquisition feeding, engorged larvae and nymphs molted to nymphs and adults, respectively, which were shown to be infected (confirming transstadial perpetuation), and were able to transmit both strains of R. rickettsii to uninfected animals, as demonstrated by clinical, serological, and molecular analyses. However, the larval, nymphal, and adult stages of A. dubitatum showed to be only partially susceptible to R. rickettsii infection, since in all cases, only part of the ticks became infected by this agent, after being exposed to rickettsemic animals. While transovarial transmission of R. rickettsii was inefficient in the A. dubitatum engorged females of the present study, 100% of these females passed R. bellii transovarially. Because it has been reported that a primary infection by a Rickettsia species would preclude transovarial transmission of a second Rickettsia species, it is likely that the ineffectiveness of A. dubitatum to perpetuate R. rickettsii by transovarial transmission was related to its primary infection by R. bellii; however, it could also be related to unknown factors inherent to A. dubitatum. The relevance of A. dubitatum as a natural vector of R. rickettsii to humans or animals is discussed.
Darke, Sabina; Marschang, Rachel E; Hetzel, Udo; Reinacher, Manfred
Orthoreoviruses have been associated with disease in reptiles, but have not previously been isolated from snakes with inclusion body disease (IBD). An orthoreovirus was isolated from a Boa constrictor diagnosed with IBD and then used to conduct a transmission study to determine the clinical importance of this virus. For the transmission study, 10 juvenile boas were experimentally infected with the isolated orthoreovirus and compared to 5 sham-infected control animals. Orthoreovirus was reisolated for a period of 18 wk after infection and weight gain was reduced in infected snakes. Histological examination showed a mild hepatitis in three of four virologically positive snakes up to 12 wk after infection. Results indicated that the orthoreovirus was moderately pathogenic, but, no evidence was found to indicate that it was the causal agent of IBD. In the light of the discovery of Arenaviruses in some snakes with IBD, it was proposed that orthoreoviruses may play a role in synergistic infection.
English, R V; Nelson, P; Johnson, C M; Nasisse, M; Tompkins, W A; Tompkins, M B
Cats naturally infected with feline immunodeficiency virus (FIV) develop an AIDS-like syndrome whereas experimentally infected cats do not. To investigate the role of cofactors in the development of this disease in cats, 7 specific pathogen-free (SPF) and 12 random-source (RS) cats were infected with FIV. Over 4 years, infected cats developed similar phenotypic and functional immune abnormalities characterized by early and chronic inversion of CD4+:CD8+ cell ratios and significantly decreased mitogen responses compared with controls. Beginning 18-24 months after infection, 10 RS cats developed chronic clinical disease typical of feline AIDS, including stomatitis and recurrent upper respiratory disease; 4 SPF cats also developed chronic clinical disease, 2 with neurologic disease and 2 with B cell lymphomas. Thus, immunologic background is important in the type of disease that develops in cats infected with FIV, and FIV represents a promising animal model for studying the immunopathogenesis of AIDS in humans.
s.l., An. aquasalis and An. nuneztovari s.l. had higher infection rates than An. darlingi. Conclusion All field-collected Anopheles species, as well as colonized An. aquasalis are susceptible to experimental P. vivax infections by membrane feeding assays. Anopheles darlingi, An. albitarsis s.l. and An. aquasalis are very susceptible to P. vivax infection. However, colonized An. aquasalis mosquitoes showed the higher infection intensity represented by infection rate and oocyst numbers. This study is the first to characterize experimental development of Plasmodium infections in Amazon Anopheles vectors and also to endorse that P. vivax infection of colonized An. aquasalis is a feasible laboratory model. PMID:24359307
Abdel-Hafeez, Ekhlas H; Ahmad, Azza K; Abdelgelil, Noha H; Abdellatif, Manal Z M; Kamal, Amany M; Hassanin, Kamel M A; Abdel-Razik, Abdel-Razik H; Abdel-Raheem, Ehab M
Blastocystis spp., one of the most common parasites colonizing the human intestine, is an extracellular, luminal protozoan with controversial pathogenesis. The host's immune response against Blastocystis spp. infection has also not been defined yet. Therefore, this research aimed to assess the potential pathogenicity of this parasite and its ability to modulate the immune response in experimental infected immunocompetent and immunosuppresed mice. These results demonstrated that the infected immunosuppressed mice were more affected than infected immunocompetent mice. Histopathological examination of the small intestine in the infected immunosuppressed mice showed that Blastocystis spp. infiltrated all the layers. Moreover, the epithelia showed exfoliation and inflammatory cell infiltration in submucosa compared to that of the infected immunocompetent mice. As well, examination of the large intestine of the infected immunosuppressed group showed severe goblet cell hyperplasia. Blastocystis spp. infiltrated all the large intestine layers compared to that of the infected immunocompetent group. Furthermore, there was a significant upregulation of the expression of proinflammatory cytokines: interleukin 12 (IL-12) and tumor necrosis factor alpha (TNF-α) in the infected immunosuppressed mice compared to that of the infected immunocompetent ones (p ≤ 0.004 and p ≤ 0.002, respectively). However, the expression of anti-inflammatory cytokines (IL-4 and IL-10) was significantly downregulated in the infected immunosuppressed group compared to that of the infected immunocompetent group one at 10 days postinfection (p ≤ 0.002 and p ≤ 0.001, respectively). The results of this study revealed that Blastocystis spp. affected the production of pro- and anti-inflammatory cytokines in both groups of mice compared to healthy normal (naive) group. Additionally, these data showed that there was a significant upregulation (p ≤ 0.005) of the locally
Oosterik, Leon H; Tuntufye, Huruma N; Tsonos, Jessica; Luyten, Tom; Noppen, Sam; Liekens, Sandra; Lavigne, Rob; Butaye, Patrick; Goddeeris, Bruno M
Avian pathogenic Escherichia coli (APEC) are responsible for significant economic losses in the poultry industry. In this study, a model for investigating the pathogenesis of APEC infections was established. APEC strain CH2 (O78) was marked with the luciferase operon (luxCDABE) using a Tn7 transposon and tissues of experimentally infected chickens were analysed for a correlation between the bioluminescent signal and the number of bacteria. Transposition of the lux operon into the chromosome of the APEC isolate did not affect sensitivity to lytic bacteriophages and there was no effect on virulence in an intratracheal infection model in 1-day-old chicks, although results with a subcutaneous infection model were inconclusive. A correlation between the number of bacteria and the luminescent signal was found in liquid medium, as well as in homogenised heart, liver, spleen and lung of 4-week-old experimentally infected chickens. This study showed that lux could be used for identification of the infecting strain after experimental infection with APEC in poultry.
Calhoun, Dana M.; Schaffer, Paula A.; Gregory, Jacklyn R.; Hardy, Katherine M.; Johnson, Pieter T. J.
Infections by the digenetic trematode, Ribeiroia ondatrae, cause severe limb malformations in many North American amphibians. Ribeiroia ondatrae also infects fishes as second intermediate hosts, but less is known about the pathology and immune responses initiated in infected fish, even though reports of infected fish date back to early 1900s. To this end, we experimentally exposed juvenile Bluegills Lepomis macrochirus to three doses of R. ondatrae cercariae and monitored the pathology, parasite infection success, and humoral responses over 648 h. All exposed fish became infected with metacercariae, and the average infection load increased with exposure dose. Histologically, infection was associated with acute hemorrhages in the lateral line and local dermis at 36 h, followed by progressive granulomatous inflammation that led to the destruction of encysted metacercariae. Correspondingly, over the course of 648 h we observed an 85% decline in average infection load among hosts, reflecting the host’s clearance of the parasite. Infection was not associated with changes in fish growth or survival, but did correlate with leukocytosis and neutrophilia in circulating host blood. Understanding the physiological responses of R. ondatrae in Bluegill will help to clarify the ecological effects of this parasite and provide a foundation for subsequent comparisons into its effects on behavior, individual health, and population dynamics of Bluegill. PMID:26587684
Mortatti, R C; Maia, L C; de Oliveira, A V; Munk, M E
The immunopathology of Chagas' disease was studied in the experimental model of chronic infection in C57BL/10JT or mice. Sublethal infection with Trypanosoma cruzi, Y strain, induced specific antibodies and a delayed hypersensitivity response to parasite antigens. Mice developed chronic chagasic myocarditis but not skeletal muscle myositis. Binding of T cells to infected heart tissue was investigated during short-term cocultivation of lymphocytes with heart cryostat sections. T cells from infected mice and from normal controls bound equally to myocardium and liver sections from both infected and normal mice. A search in depth was attempted with cells heavily tagged with 99mTc. Labeled T cells from chagasic mice bound to both normal and infected myocardium slices. 99mTc-labeled T cells from controls gave the same binding values. Glass-adherent spleen cells behaved identically to T cells. Prior treatment of the tissue with serum from chronically infected mice did not increase the number of binding cells. Peritoneal macrophages tagged with 99mTc-sulfur colloid also bound to infected myocardium slices. The binding of macrophages was not changed by pretreatment of infected tissue with anti-T, cruzi antibodies. In short, this work did not detect any population of T cells or macrophages which could bind specifically to infected heart tissue to initiate an autoreactive process. Images PMID:2228230
Quirk, Travis; Dubey, J P
Twenty-three striped skunks (Mephitis mephitis) without demonstrable antibodies in 1:25 serum dilution in the modified agglutination test (MAT) were fed sporulated Toxoplasma gondii oocysts (9 skunks) or tissue cysts (10 skunks), and 4 skunks (controls) were not fed T. gondii. Skunks were bled before feeding T. gondii, 10 and 23- 25 days postinoculation (PI). All 9 seronegative skunks fed oocysts died of acute toxoplasmosis between 7 and 19 days PI; T. gondii tachyzoites were found in histological sections of many tissues. One of the 10 skunks fed tissue cysts and 1 of the 4 controls also died of acute toxoplasmosis days 19 and 20 PI; these animals probably became infected by ingestion of unexcysted oocysts passed in feces of skunks fed oocysts that were housed in the same room that skunks fed tissue cysts were housed. The remaining 9 skunks fed tissue cysts and the 3 controls developed only a mild illness and were killed in good health on days 23-25 PI. Antibodies to T. gondii were not found in 1:25 serum dilution of any of the 19 of 23 skunks that were alive on day 10 PI; 12 of 13 skunks had antibodies (MAT 1:80 or higher) on the day they were killed. Antibodies were not found in 1 skunk. Results indicate that skunks can develop IgG antibodies to T. gondii within 3 wk PI, and primary toxoplasmosis can be fatal in skunks.
Fan, P C; Lin, C Y; Chung, W C
In the present study, six 34-44-day-old Small-Ear-Miniature pigs and one 14-day-old Holstein calf were each fed 10,000 Philippine Taenia eggs and sacrificed 27-43 days after inoculation. The infection rate was 100% for both pigs and calf with cysticerci recovery rates of 11 and 6%, respectively. A total of 6431 cysticerci were recovered only from the livers of the six pigs and 597 only from the liver of the calf; more occurred in the parenchyma (pigs 75%, calf 83%) than on the surface (pigs 25%, calf 17%). Mature cysticerci were found in four of the six pigs. A total of 317 cysticerci recovered from the pig livers were mature and the rest were either immature (926), degenerate or calcified (5188). All 597 cysticerci recovered from the liver of the calf were degenerate or calcified. Measurements of length, width, diameter of protoscolex, rostellum, and sucker and hooklet pattern indicated that Philippine Taenia is very similar to Taenia from Taiwan, Korea, Indonesia and Thailand and very different from classical T. saginata and T. solium.
Cammock, Cheryl E.; Halnon, Nancy J.; Skoczylas, Jill; Blanchard, James; Bohm, Rudolf; Miller, Christopher J.; Lai, Chi; Krogstad, Paul A.
Coxsackievirus B (CVB) infection is a common cause of acute viral myocarditis. The clinical presentation of myocarditis caused by this enterovirus is highly variable, ranging from mildly symptoms to complete hemodynamic collapse. These variations in initial symptoms and in the immediate and long term outcomes of this disease have impeded development of effective treatment strategies. Nine cynomolgus monkeys were inoculated with myocarditic strains of CVB. Virological studies performed up to 28 days post-inoculation demonstrated the development of neutralizing antibody in all animals, and the presence of CVB in plasma. High dose intravenous inoculation (n = 2) resulted in severe disseminated disease, while low dose intravenous (n = 6) or oral infection (1 animal) resulted in clinically unapparent infection. Transient, minor, echocardiographic abnormalities were noted in several animals, but no animals displayed signs of significant acute cardiac failure. Although viremia rapidly resolved, signs of myocardial inflammation and injury were observed in all animals at the time of necropsy, and CVB was detected in postmortem myocardial specimens up to 28 days PI. This non-human primate system replicates many features of illness in acute coxsackievirus myocarditis and demonstrates that myocardial involvement may be common in enteroviral infection; it may provide a model system for testing of treatment strategies for enteroviral infections and acute coxsackievirus myocarditis. PMID:24040287
Nemeth, Nicole; Gould, Daniel; Bowen, Richard; Komar, Nicholas
We studied the effects of natural and/or experimental infections of West Nile virus (WNV) in five raptor species from July 2002 to March 2004, including American kestrels (Falco sparverius), golden eagles (Aquila chrysaetos), red-tailed hawks (Buteo jamaicensis), barn owls (Tyto alba), and great horned owls (Bubo virginianus). Birds were infected per mosquito bite, per os, or percutaneously by needle. Many experimentally infected birds developed mosquito-infectious levels of viremia (>10(5) WNV plaque forming units per ml serum) within 5 days postinoculation (DPI), and/ or shed virus per os or per cloaca. Infection of organs 15-27 days postinoculation was infrequently detected by virus isolation from spleen, kidney, skin, heart, brain, and eye in convalescent birds. Histopathologic findings varied among species and by method of infection. The most common histopathologic lesions were subacute myocarditis and encephalitis. Several birds had a more acute, severe disease condition represented by arteritis and associated with tissue degeneration and necrosis. This study demonstrates that raptor species vary in their response to WNV infection and that several modes of exposure (e.g., oral) may result in infection. Wildlife managers should recognize that, although many WNV infections are sublethal to raptors, subacute lesions could potentially reduce viability of populations. We recommend that raptor handlers consider raptors as a potential source of WNV contamination due to oral and cloacal shedding.
Kumar, Sachin; Militino Dias, Flavia; Nayak, Baibaswata; Collins, Peter L; Samal, Siba K
Avian paramyxoviruses (APMV) are divided into nine serotypes. Newcastle disease virus (APMV-1) is the most extensively characterized, while relatively little information is available for the other APMV serotypes. In the present study, we examined the pathogenicity of two divergent strains of APMV-3, Netherlands and Wisconsin, in (i) 9-day-old embryonated chicken eggs, (ii) 1-day-old specific pathogen free (SPF) chicks and turkeys, and (iii) 2-week-old SPF chickens and turkeys. The mean death time in 9-day-old embryonated chicken eggs was 112 h for APMV-3 strain Netherlands and > 168 h for strain Wisconsin. The intracerebral pathogenicity index in 1-day-old chicks for strain Netherlands was 0.39 and for strain Wisconsin was zero. Thus, both strains are lentogenic. Both the strains replicated well in brain tissue when inoculated intracerebrally in 1-day-old SPF chicks, but without causing death. Mild respiratory disease signs were observed in 1-day-old chickens and turkeys when inoculated through oculonasal route with either strain. There were no overt signs of illness in 2-weeks-old chickens and turkeys by either strain, although all the birds seroconverted after infection. The viruses were isolated predominantly from brain, lungs, spleens, trachea, pancreas and kidney. Immunohistochemistry studies also showed the presence of large amount of viral antigens in both epithelial and sub-epithelial lining of respiratory and alimentary tracts. Our result suggests systemic spread of APMV-3 even though the viral fusion glycoprotein does not contain the canonical furin proteases cleavage site. Furthermore, there was little or no disease despite systemic viral spread and abundant viral replication in all the tissues tested.
Klix, A.; Fischer, U.; Gehre, D.; Kleizer, G.; Raj, P.; Rovni, I.; Ruecker, Tom
We are investigating methods for neutron flux measurement in the ITER TBM. In particular we have tested sets of activation materials leading to induced gamma activities with short half-lives of the order of tens of seconds up to minutes and standard activation materials. Packages of activation foils have been irradiated with the intense neutron generator of Technical University of Dresden in a pure DT neutron field as well as in a neutronics mock-up of the European ITER HCLL TBM. An important aim was to check whether the gamma activity induced in the activation foils in these packages could be measured simultaneously. It was indeed possible to identify gamma lines of interest in gamma-ray measurements immediately after extraction from the irradiation.
Galindo-Cardiel, I; Busquets, N; Velarde, R; Abad, F X; Solanes, D; Rivas, R; Valle, R; Brun, A; Domingo, M
Lymphoplasmacytic endotheliitis and anterior uveitis was diagnosed in four lambs infected experimentally with field isolates of Rift Valley fever virus (RVFV). Formalin-fixed and paraffin wax-embedded tissue from these animals was investigated by histopathology and quantitative real time reverse transcriptase polymerase chain reaction. To our knowledge, this is the first pathological description of this ocular manifestation of RVFV infection in ruminants, although these lesions have been described in man.
Bluetongue (BT) is an insect-transmitted, economically important disease of domestic and wild ruminants. Although only five of the 26 reported bluetongue virus (BTV) serotypes are considered endemic to the USA, 10 exotic serotypes have been isolated primarily in the southeastern region of the count...
Gharbi, M; Latrach, R; Sassi, L; Darghouth, M A
We have evaluated a new simple technique using whole blood from experimentally infected cattle for the isolation and cultivation of Theileria annulata. The study was carried out on 20 Holstein-Frisian bovines that had been experimentally infected with a virulent lethal dose of Theileria annulata. This technique has been compared to the classical peripheral blood monocyte isolation with Ficoll carried out on 22 experimentally infected Holstein-Friesian calves. The effectiveness of the reference technique was estimated to 86.4%, whilst the effectiveness of the new technique was 100%. Moreover, this new technique leads to time and money saving estimated to € 3.06 per sample. It decreases the contamination risks by reducing the steps of sample manipulation.
Gardner, D E
The success or failure of the respiratory system to defend itself against airborne infectious agents largely depends upon the efficiency of the pulmonary defenses to maintain sterility and to dispose of unwanted substances. Both specific and nonspecific host defenses cooperate in the removal and inactivation of such agents. Several studies have shown that these defenses are vulnerable to a wide range of environmental agents and that there is a good relationship between exposure to pollutant and the impaired resistance to pulmonary disease. There are numerous immunological, biochemical and physiological techniques that are routinely used to identify and to characterize individual impairments of these defenses. Based on these effects, various hypotheses are proposed as to what health consequences could be expected from these effects. The ultimate test is whether the host, with its compromised defense mechanisms, is still capable of sustaining the total injury and continuing to defend itself against opportunistic pathogens. This paper describes the use of an experimental airborne infectious disease model capable of predicting subtle changes in host defenses at concentrations below which there are any other overt toxicological effects. Such sensitivity is possible because the model measure not just a single "health" parameter, but instead is capable of reflecting the total responses caused by the test chemical. Images FIGURE 3. PMID:7060549
Hope, Vivian D; Palmateer, Norah; Wiessing, Lucas; Marongiu, Andrea; White, Joanne; Ncube, Fortune; Goldberg, David
The recent anthrax outbreak among injecting drug users (IDUs) in Europe has highlighted an ongoing problem with severe illness resulting from spore-forming bacteria in IDUs. We collated the numbers of cases of 4 bacterial illnesses (botulism, tetanus, Clostridium novyi, and anthrax) in European IDUs for 2000 to 2009 and calculated population rates. Six countries reported 367 cases; rates varied from 0.03 to 7.54 per million people. Most cases (92%) were reported from 3 neighboring countries: Ireland, Norway, and the United Kingdom. This geographic variation needs investigation.
Da Silva, Aleksandro S; Fausto, Guilherme C; Grando, Thirssa H; Cadore, Carlos A; Pimentel, Victor C; Jaques, Jeandre A; Schetinger, Maria R C; Monteiro, Silvia G; Leal, Marta L R
The aim of this study was to evaluate adenosine deaminase (E-ADA) activity in sera of lambs experimentally infected with Haemonchus contortus. We used 12 lambs divided into 2 groups; Group A had 5 healthy, non-infected animals (control) and Group B had 7 healthy animals infected with H. contortus . Lambs were infected orally with 500 larvae (L3) per animal every 2 days, for a period of 20 days, and later the infection was confirmed by examination of feces (eggs per gram [EPG] via fecal egg count). Blood collection was performed at days 0, 20, 40, 60, and 80 post-infection (PI) for analysis of E-ADA activity. Animals in Group A showed negative EPG throughout the experiment unlike those from Group B that had elevated EPG counts. E-ADA activity was reduced in the serum of animals infected with H. contortus when compared to non-infected controls at days 20, 40, 60, and 80 PI. Therefore, it is concluded that infection with H. contortus influences the E-ADA activity in lambs.
Watts, D M; Tammariello, R F; Dalrymple, J M; Eldridge, B F; Russell, P K; Top, F H
Experimental studies were conducted to assess the susceptibility of white-tailed deer (Odocoileus virginianus), gray squirrels (Sciurus carolinensis), and cottontail rabbits (Sylvilagus floridanus) to Jamestown Canyon (JC) and/or Keystone (KEY) virus infection. Viremia occurred in 5 of 6 deer inoculated with JC virus; however, all deer developed KEY virus neutralizing antibody. Based on the observation that antibody elicited by primary infection of deer with either KEY or JC virus exhibited partial heterologous neutralization in vitro, cross-challenge experiments were performed in these animals. Keystone virus failed to infect deer 30 days post primary JC virus infection; however, deer became infected when challenged with KEY virus 80 days after the initial JC virus infection as indicated by a substantial increase in antibody titer. Similarly, JC virus failed to produce viremia in immune animals infected with KEY virus 80 days previously, although 2 of the 3 animals challenged had serological evidence of infection. Three field-collected cottontail rabbits with no evidence of KEY antibody were readily susceptible to KEY virus infection and developed viremias of 1-4 days' duration; rabbits with KEY virus antibody did not develop viremia upon KEY virus challenge. Eight antibody-negative field-collected gray squirrels became viremic following injection with KEY virus; however, a comparable group of squirrels did not become viremic when injected with JC virus.
Nettle, Daniel; Andrews, Clare; Reichert, Sophie; Bedford, Tom; Kolenda, Claire; Parker, Craig; Martin-Ruiz, Carmen; Monaghan, Pat; Bateson, Melissa
Early-life adversity is associated with accelerated cellular ageing during development and increased inflammation during adulthood. However, human studies can only establish correlation, not causation, and existing experimental animal approaches alter multiple components of early-life adversity simultaneously. We developed a novel hand-rearing paradigm in European starling nestlings (Sturnus vulgaris), in which we separately manipulated nutritional shortfall and begging effort for a period of 10 days. The experimental treatments accelerated erythrocyte telomere attrition and increased DNA damage measured in the juvenile period. For telomere attrition, amount of food and begging effort exerted additive effects. Only the combination of low food amount and high begging effort increased DNA damage. We then measured two markers of inflammation, high-sensitivity C-reactive protein and interleukin-6, when the birds were adults. The experimental treatments affected both inflammatory markers, though the patterns were complex and different for each marker. The effect of the experimental treatments on adult interleukin-6 was partially mediated by increased juvenile DNA damage. Our results show that both nutritional input and begging effort in the nestling period affect cellular ageing and adult inflammation in the starling. However, the pattern of effects is different for different biomarkers measured at different time points. PMID:28094324
Lewis, S. Rochelle; Ellison, Siobhan P.; Dascanio, John J.; Lindsay, David S.; Gogal, Robert M.; Werre, Stephen R.; Surendran, Naveen; Breen, Meghan E.; Heid, Bettina M.; Andrews, Frank M.; Buechner-Maxwell, Virginia A.; Witonsky, Sharon G.
Sarcocystis neurona is the most common cause of Equine Protozoal Myeloencephalitis (EPM), affecting 0.5–1% horses in the United States during their lifetimes. The objective of this study was to evaluate the equine immune responses in an experimentally induced Sarcocystis neurona infection model. Neurologic parameters were recorded prior to and throughout the 70-day study by blinded investigators. Recombinant SnSAG1 ELISA for serum and CSF were used to confirm and track disease progression. All experimentally infected horses displayed neurologic signs after infection. Neutrophils, monocytes, and lymphocytes from infected horses displayed significantly delayed apoptosis at some time points. Cell proliferation was significantly increased in S. neurona-infected horses when stimulated nonspecifically with PMA/I but significantly decreased when stimulated with S. neurona compared to controls. Collectively, our results suggest that horses experimentally infected with S. neurona manifest impaired antigen specific response to S. neurona, which could be a function of altered antigen presentation, lack of antigen recognition, or both. PMID:26464923
Lewis, S Rochelle; Ellison, Siobhan P; Dascanio, John J; Lindsay, David S; Gogal, Robert M; Werre, Stephen R; Surendran, Naveen; Breen, Meghan E; Heid, Bettina M; Andrews, Frank M; Buechner-Maxwell, Virginia A; Witonsky, Sharon G
Sarcocystis neurona is the most common cause of Equine Protozoal Myeloencephalitis (EPM), affecting 0.5-1% horses in the United States during their lifetimes. The objective of this study was to evaluate the equine immune responses in an experimentally induced Sarcocystis neurona infection model. Neurologic parameters were recorded prior to and throughout the 70-day study by blinded investigators. Recombinant SnSAG1 ELISA for serum and CSF were used to confirm and track disease progression. All experimentally infected horses displayed neurologic signs after infection. Neutrophils, monocytes, and lymphocytes from infected horses displayed significantly delayed apoptosis at some time points. Cell proliferation was significantly increased in S. neurona-infected horses when stimulated nonspecifically with PMA/I but significantly decreased when stimulated with S. neurona compared to controls. Collectively, our results suggest that horses experimentally infected with S. neurona manifest impaired antigen specific response to S. neurona, which could be a function of altered antigen presentation, lack of antigen recognition, or both.
Rehfeld, Izabelle S; Guedes, Maria Isabel M C; Matos, Ana Carolina D; de Oliveira, Tércia M L; Rivetti, Anselmo V; Moura, Ana Carolina J; Paes, Paulo Ricardo O; do Lago, Luiz Alberto; Kroon, Erna G; Lobato, Zélia Inês P
Vaccinia virus (VACV) is the etiological agent of bovine vaccinia (BV), an important zoonosis that affects dairy cattle. There are many aspects of the disease that remain unknown, and aiming to answer some of these questions, the clinical, hematological, and biochemical parameters of VACV experimentally infected cows were evaluated. In the first part of the study, lactating cows were infected with VACV-GP2 strain. In the second part, animals previously infected with VACV-GP2 were divided into two treatment groups: Group 1, immunosuppressed cows; and Group 2, re-infected cows. In this study, BV could be experimentally reproduced, with similar lesions as observed in natural infections. Moreover, a short incubation period and local lymphadenopathy were also observed. VACV could be detected by PCR and isolated from scabs taken from teat lesions of all inoculated and re-inoculated animals. Lymphocytosis and neutrophilia were observed in all animals from the first part of the experiment, and lymphopenia and relative neutrophilia were observed in the immunosuppressed animals. Detection of viral DNA in oral mucosa lesions suggests that viral reactivation might occur in immunosuppressed animals. Moreover, clinical disease with teat lesions may occur in previously VACV-infected cows under the experimental conditions of the present study.
Bansal, G C; Sharma, N N
The antitheilerial activity of buparvaquone (BW 720C) was evaluated in experimentally induced Theileria annulata infections in cross-bred male calves. T. annulata infections were induced by injecting a suspension of infected ground tick tissue suspension (GUTTS) equivalent to two ticks subcutaneously into each calf. Buparvaquone at a dose of 2.5 mg kg-1 body weight was given as a single injection (intramuscularly) on Day 0 (Group 1), Day 8 (Group 2) and Day 12 (Group 3) post-infection. The animals in Groups 4 and 5 were untreated and challenged controls, respectively. All of the recovered animals from Groups 1-4 were challenged with a lethal dose of T. annulata at 6 weeks post-infection. The immunized animals were resistant to the homologous challenge, which killed three of four control animals (Group 5); the controls showed typical antemortem and post-mortem lesions of theileriosis.
Li, Jian-Ping; Liao, Yun; Zhang, Ying; Wang, Jing-Jing; Wang, Li-Chun; Feng, Kai; Li, Qi-Han; Liu, Long-Ding
Coxsackie virus A16 (CA16) is commonly recognized as one of the main human pathogens of hand-foot-mouth disease (HFMD). The clinical manifestations of HFMD include vesicles of hand, foot and mouth in young children and severe inflammatory CNS lesions. In this study, experimentally CA16 infected tree shrews (Tupaia belangeri) were used to investigate CA16 pathogenesis. The results showed that both the body temperature and the percentages of blood neutrophilic granulocytes / monocytes of CA16 infected tree shrews increased at 4-7 days post infection. Dynamic distributions of CA16 in different tissues and stools were found at different infection stages. Moreover, the pathological changes in CNS and other organs were also observed. These findings indicate that tree shrews can be used as a viable animal model to study CA16 infection.
LI, Jian-Ping; LIAO, Yun; ZHANG, Ying; WANG, Jing-Jing; WANG, Li-Chun; FENG, Kai; LI, Qi-Han; LIU, Long-Ding
Coxsackie virus A16 (CA16) is commonly recognized as one of the main human pathogens of hand-foot-mouth disease (HFMD). The clinical manifestations of HFMD include vesicles of hand, foot and mouth in young children and severe inflammatory CNS lesions. In this study, experimentally CA16 infected tree shrews (Tupaia belangeri) were used to investigate CA16 pathogenesis. The results showed that both the body temperature and the percentages of blood neutrophilic granulocytes / monocytes of CA16 infected tree shrews increased at 4-7 days post infection. Dynamic distributions of CA16 in different tissues and stools were found at different infection stages. Moreover, the pathological changes in CNS and other organs were also observed. These findings indicate that tree shrews can be used as a viable animal model to study CA16 infection. PMID:25465084
Wheeler, Sarah S.; Langevin, Stanley A.; Brault, Aaron C.; Woods, Leslie; Carroll, Brian D.; Reisen, William K.
To determine whether West Nile virus (WNV) persistent infection in avian hosts may potentially serve as an overwintering mechanism, House Sparrows and House Finches, experimentally and naturally infected with several strains of WNV, and two naturally infected Western Scrub-Jays were held in mosquito-proof outdoor aviaries from 2007–March 2008. Overall, 94% (n = 36) of House Sparrows, 100% (n = 14) of House Finches and 2 Western Scrub-Jays remained WNV antibody positive. When combined by species, 37% of the House Sparrows, 50% of the House Finches, and 2 Western Scrub-Jays were WNV RNA positive at necropsy, up to 36 weeks post-infection. Infectious WNV was not detected. Our study supports the hypothesis that some avian hosts support the long-term persistence of WNV RNA, but it remains unresolved whether these infections relapse to restart an avian-arthropod transmission cycle and thereby serve as an overwintering mechanism for WNV. PMID:22826479
Atkins, Helen S; Spencer, Stephen; Brew, Simon D; Jenner, Dominic C; Sefton, Armine M; MacMillan, Alastair P; Brooks, Timothy J G; Simpson, Andrew J H
The prophylactic potential of the azalide azithromycin as well as the fluoroquinolones trovafloxacin and grepafloxacin was assessed for the control of infection with Brucella melitensis in an experimental mouse model, determined by reduction in splenic bacterial burden. Trovafloxacin showed limited protective efficacy when administered 2h following a low-dose B. melitensis challenge, whereas grepafloxacin was ineffective. In comparison, azithromycin provided significant control of infection both following low- and high-dose challenges. Overall, the data confirm the potential utility of azithromycin in the prophylaxis of brucellosis and suggest that neither trovafloxacin nor grepafloxacin would likely be valuable for post-exposure prophylaxis of Brucella infection.
Jarvi, Susan I.; Schultz, Jeffrey J.; Atkinson, Carter T.
Several polymerase chain reaction (PCR)-based methods have recently been developed for diagnosing malarial infections in both birds and reptiles, but a critical evaluation of their sensitivity in experimentally-infected hosts has not been done. This study compares the sensitivity of several PCR-based methods for diagnosing avian malaria (Plasmodium relictum) in captive Hawaiian honeycreepers using microscopy and a recently developed immunoblotting technique. Sequential blood samples were collected over periods of up to 4.4 yr after experimental infection and rechallenge to determine both the duration and detectability of chronic infections. Two new nested PCR approaches for detecting circulating parasites based on P. relictum 18S rRNA genes and the thrombospondin-related anonymous protein (TRAP) gene are described. The blood smear and the PCR tests were less sensitive than serological methods for detecting chronic malarial infections. Individually, none of the diagnostic methods was 100% accurate in detecting subpatent infections, although serological methods were significantly more sensitive (97%) than either nested PCR (61–84%) or microscopy (27%). Circulating parasites in chronically infected birds either disappear completely from circulation or to drop to intensities below detectability by nested PCR. Thus, the use of PCR as a sole means of detection of circulating parasites may significantly underestimate true prevalence.
Gonzalez, María Inés; Sanjuan, Norberto A
Herpes simplex virus type 1 is one of the most frequent causes of oral infection in humans, especially during early childhood. Several experimental models have been developed to study the pathogenesis of this virus but all of them employed adult animals. In this work, we developed an experimental model that uses mice younger than 4 days old, to more closely resemble human infection. Mice were infected subcutaneously with the prototype strain McIntyre of Herpes simplex-1, and the progression of infection was studied by immunoperoxidase. All animals died within 24-72 h post-infection, while viral antigens were found in the oral epithelium, nerves and brain. The most striking result was the finding of viral antigens in the nucleus and cytoplasm of cells belonging to striated muscles. Organotypic cultures of striated muscles were performed, and viral replication was observed in them by immunocytochemistry, electron microscopy and viral isolation. We conclude that the infection of striated muscles is present from the onset of oral infection and, eventually, could explain some clinical observations in humans.
Le Goff, L; Hubert, B; Favennec, L; Villena, I; Ballet, J J; Agoulon, A; Orange, N; Gargala, G
Cryptosporidium spp., a significant cause of foodborne infection, have been shown to be resistant to most chemical food disinfectant agents and infective for weeks in irrigation waters and stored fresh vegetal produce. Pulsed UV light (PL) has the potential to inactivate Cryptosporidium spp. on surfaces of raw or minimally processed foods or both. The present study aimed to evaluate the efficacy of PL on viability and in vivo infectivity of Cryptosporidium parvum oocysts present on raspberries, a known source of transmission to humans of oocyst-forming apicomplexan pathogens. The skin of each of 20 raspberries was experimentally inoculated with five 10-μl spots of an oocyst suspension containing 6 × 10(7) oocysts per ml (Nouzilly isolate). Raspberries were irradiated by PL flashes (4 J/cm(2) of total fluence). This dose did not affect colorimetric or organoleptic characteristics of fruits. After immunomagnetic separation from raspberries, oocysts were bleached and administered orally to neonatal suckling mice. Seven days after infection, mice were euthanized, and the number of oocysts in the entire small intestine was individually assessed by immunofluorescence flow cytometry. Three of 12 and 12 of 12 inoculated mice that received 10 and 100 oocysts isolated from nonirradiated raspberries, respectively, were found infected. Four of 12 and 2 of 12 inoculated mice that received 10(3) and 10(4) oocysts from irradiated raspberries, respectively, were found infected. Oocyst counts were lower in animals inoculated with 10(3) and 10(4) oocysts from irradiated raspberries (92 ± 144 and 38 ± 82, respectively) than in animals infected with 100 oocysts from nonirradiated raspberries (35,785 ± 66,221, P = 0.008). PL irradiation achieved oocyst reductions of 2 and 3 log for an inoculum of 10(3) and 10(4) oocysts, respectively. The present pilot-scale evaluation suggests that PL is an effective mode of decontamination for raspberries and prompts further applicability
Saville, W J A; Dubey, J P; Oglesbee, M J; Sofaly, C D; Marsh, A E; Elitsur, E; Vianna, M C; Lindsay, D S; Reed, S M
Sarcocystis neurona and Sarcocystis fayeri infections are common in horses in the Americas. Their antemortem diagnosis is important because the former causes a neurological disorder in horses, whereas the latter is considered nonpathogenic. There is a concern that equine antibodies to S. fayeri might react with S. neurona antigens in diagnostic tests. In this study, 4 ponies without demonstrable serum antibodies to S. neurona by Western immunoblot were used. Three ponies were fed 1 x 10(5) to 1 x 10(7) sporocysts of S. fayeri obtained from dogs that were fed naturally infected horse muscles. All ponies remained asymptomatic until the termination of the experiment, day 79 postinoculation (PI). All serum samples collected were negative for antibodies to S. neurona using the Western blot at the initial screening, just before inoculation with S. fayeri (day 2) and weekly until day 79 PI. Cerebrospinal fluid samples from each pony were negative for S. neurona antibodies. Using the S. neurona agglutination test, antibodies to S. neurona were not detected in 1:25 dilution of sera from any samples, except that from pony no. 4 on day 28; this pony had received 1 X 10(7) sporocysts. Using indirect immunofluorescence antibody tests (IFATs), 7 serum samples were found to be positive for S. neurona antibodies from 1:25 to 1:400 dilutions. Sarcocystis fayeri sarcocysts were found in striated muscles of all inoculated ponies, with heaviest infections in the tongue. All sarcocysts examined histologically appeared to contain only microcytes. Ultrastructurally, S. fayeri sarcocysts could be differentiated from S. neurona sarcocysts by the microtubules (mt) in villar protrusions on sarcocyst walls; in S. fayeri the mt extended from the villar tips to the pellicle of zoites, whereas in S. neurona the mt were restricted to the middle of the cyst wall. Results indicate that horses with S. fayeri infections may be misdiagnosed as being S. neurona infected using IFAT, and further research
El-Sayed, Nagwa Mostafa; Fathy, Ghada Mahmoud; Abdel-Rahman, Sara Abdel-Rahman; El-Shafei, Mahmoud Abdel-Atei
The purpose of this study was to determine cytokine patterns in experimental schistosomiasis mansoni infected mice treated with silymarin. The study was conducted upon 100 mice that were divided into five groups; 20 each: uninfected control group, Schistosoma mansoni infected untreated mice (infected control), infected mice treated with praziquantel (PZQ), infected mice treated with silymarin and infected mice treated with both praziquantel and silymarin. 10 mice from each group were sacrificed at 10th and 18th weeks post infection respectively. Histopathological investigations were performed. Liver sections were stained with hematoxylin-eosin and Masson's trichrome stain to evaluate changes of granuloma sizes and numbers. Serum levels of the cytokines (TNF-α, IFN-γ, IL-4 and TGF-β1) were assessed in the sera of all groups by immunoassay. The measured levels of cytokines (IFN-γ, IL-4, TNF-α, TGF-β1) were found to be significantly increased in infected mice compared to normal control. At the same time, treated groups with silymarin alone or combined with PZQ showed significant decrease in IL-4, TNF-α and TGF-β1 levels compared to infected control. On the other hand, there was a significant increase in IFN-γ level observed in all treated groups compared to infected control. In addition, the histopathological examination of the liver in the group treated with PZQ showed a reduction in the number of livers eggs granuloma at all periods of sacrification compared with the infected untreated group. However, there was more decrease in granulomas diameter in both silymarin treated group or combined with PZQ at all periods of sacrification when compared to infected untreated group. In conclusion; treatment with silymarin combined with PZQ in murine schistosomiasis could reduce hepatic fibrosis by their action on the production of pro-inflammatory cytokines.
Joachim, Anja; Schwarz, Lukas; Hinney, Barbara; Ruttkowski, Bärbel; Vogl, Claus; Mundt, Hans-Christian
For reliable predictions of clinical and parasitological outcome of experimental infections with parasites, different models must be evaluated for possible influences of infection time point, infection dose and host-specific parameters such as breed or litter size. To address these issues for Cystoisospora (syn. Isospora) suis, the causative agent of porcine neonatal coccidiosis, 181 piglets from 90 litters (hybrid crosses of different breeds) were included in a retrospective study to evaluate differences in time point and dose of infection in four different experimental models ((1) 1,500 oocysts on the 4th day of life, d.o.l.; (2) 1,000 oocysts, 4th d.o.l.; (3) 1,000 oocysts, 1st d.o.l.; (4) 5,000 oocysts, 4th d.o.l.). The target variables body weight gain, faecal consistency and oocyst excretion were evaluated during the acute phase of infection (5-10 days post infection), and the influences of the dependent variables breed or litter size were estimated. Despite differences in the time course of excretion and faecal consistency, neither the average amount of excretion nor the average faecal consistency differed among models, breeds or litters of different size. High individual variability was seen in all four models as described earlier for higher infection doses. When infections on the 1st vs. 4th day of life were compared, no differences in averages could be found, in contrast to previous observations on the influence of age. Other, not yet defined, variables appear to have a greater impact on the outcome of infection than doses and time points in the tested range, despite the reliable outcome of infection with high excretion rates and signs of clinical disease.
Debast, S B; Bauer, M P; Kuijper, E J
In 2009 the first European Society of Clinical Microbiology and Infection (ESCMID) treatment guidance document for Clostridium difficile infection (CDI) was published. The guideline has been applied widely in clinical practice. In this document an update and review on the comparative effectiveness of the currently available treatment modalities of CDI is given, thereby providing evidence-based recommendations on this issue. A computerized literature search was carried out to investigate randomized and non-randomized trials investigating the effect of an intervention on the clinical outcome of CDI. The Grades of Recommendation Assessment, Development and Evaluation (GRADE) system was used to grade the strength of our recommendations and the quality of the evidence. The ESCMID and an international team of experts from 11 European countries supported the process. To improve clinical guidance in the treatment of CDI, recommendations are specified for various patient groups, e.g. initial non-severe disease, severe CDI, first recurrence or risk for recurrent disease, multiple recurrences and treatment of CDI when oral administration is not possible. Treatment options that are reviewed include: antibiotics, toxin-binding resins and polymers, immunotherapy, probiotics, and faecal or bacterial intestinal transplantation. Except for very mild CDI that is clearly induced by antibiotic usage antibiotic treatment is advised. The main antibiotics that are recommended are metronidazole, vancomycin and fidaxomicin. Faecal transplantation is strongly recommended for multiple recurrent CDI. In case of perforation of the colon and/or systemic inflammation and deteriorating clinical condition despite antibiotic therapy, total abdominal colectomy or diverting loop ileostomy combined with colonic lavage is recommended.
Campigotto, Gabriela; Da Silva, Aleksandro S; Volpato, Andreia; Balzan, Alexandre; Radavelli, Willian M; Soldá, Natan M; Grosskopf, Hyolanda M; Stefani, Lenita M; Bianchi, Anderson E; Monteiro, Silvia G; Tonin, Alexandre A; Weiss, Paulo H E; Miletti, Luiz C; Lopes, Sonia T A
The aims of this study were to evaluate vertical transmission of Trypanosoma evansi in sheep experimentally infected, in addition to the mammary transmission by colostrum or milk of these infected sheep to mice. Three pregnant sheep were used: one uninfected, four months pregnant (Sheep A); and two (Sheep B and C) infected intravenously by T. evansi trypomastigotes (4.6×10(6) per animal) on the third (Sheep C) and fourth (Sheep B) month of pregnancy. Both infected sheep developed low and oscillating parasitemia measured by blood smears. Hemogram was performed at seven day intervals, showing anemia, leukocytosis, and lymphocytosis on sheep B and C. Three sheep had twins, where sheep A delivered healthy lambs and both infected sheep had delivered at least one stillborn. Additionally, lambs from sheep B and C died 24 and 72 h post-partum, respectively. Before colostrum intake, four lambs from infected sheep were positives for T. evansi according to blood smear evaluation, serology (CATT/T. evansi), and PCR. Sheep colostrum and milk samples collected from the first four days post-partum were positives for T. evansi on PCR, and these samples were able to infect seven mice (out of 10) orally (n=4/5) and intraperitoneally (n=3/5). Therefore, we conclude that the vertical transmission of T. evansi occurs in pregnant sheep, in addition to a strong possibility of the transmission by colostrum and milk.
Carnicer-Pont, Dolors; de Olalla, Patricia G; Caylă, Joan A
The study goal is to identify predictors of HIV infection late detection in an European city with increased immigration, and determine the effects of HAART era in HIV infection detection. We used Barcelona city AIDS registry (1987-2006). Late testers were those diagnosed of AIDS defining illness within less than 3 months from time of testing positive for HIV infection. Independent variables were: date of birth, sex, country of origin, HIV transmission category, prison history, city district of residence, AIDS diagnostic disease and HAART era when diagnosed. The statistical methods were based on logistic regression (Odds Ratio, OR and 95% confidence interval, CI). Among the 6186 AIDS patients, 43.9% (n=2741) were late testers. Being a male (OR: 1.57, 95% CI: 1.35-1.83), either < 30 years (OR: 1.21, 95% CI: 1.06-1.38) or > 40 years (OR: 1.20, 95% CI: 1.03-1.40), with heterosexual (OR: 3.07, 95% CI: 2.59-3.63) routes of transmission or men who have sex with men (OR: 2.20, 95% CI: 1.89-2.57) and with Pneumocystis jiroveci pneumoniae (OR: 1.71, 95% CI: 1.47-2.00) or tuberculosis (OR: 1.57, 95% CI: 1.36-1.82) were all independent risk factors for being a late tester. Conversely, injecting drug use (IDU) was associated with early detection (OR: 0.36, 95% CI: 0.33-0.40). Being migrant was associated with late testing only in the univariate analysis. Individuals with the detected factors (male, having any sexual risk behaviour and being > 50 years) should be in the main focus for HIV testing to further ensure continuous decrease in the slope of late detected HIV infections overall.
Schoo, Katherina L; Aberle, Nicole; Malzahn, Arne M; Boersma, Maarten
The issues of food quality and food quantity are crucial for trophic interactions. Although most research has focussed on the primary producer-herbivore link, recent studies have shown that quality effects at the bottom of the food web propagate to higher trophic levels. Negative effects of poor food quality have almost exclusively been demonstrated at higher food quantities. Whether these negative effects have the same impact at low food availability in situations where the majority if not all of the resources are channelled into routine metabolism, is under debate. In this study a tri-trophic food chain was designed, consisting of the algae Rhodomonas salina, the copepod Acartia tonsa and freshly hatched larvae of the European lobster Homarus gammarus. The lobster larvae were presented with food of two different qualities (C:P ratios) and four different quantities to investigate the combined effects of food quality and quantity. Our results show that the quality of food has an impact on the condition of lobster larvae even at very low food quantities. Food with a lower C:P content resulted in higher condition of the lobster larvae regardless of the quantity of food. These interacting effects of food quality and food quantity can have far reaching consequences for ecosystem productivity.
Mukhin, Andrey; Palinauskas, Vaidas; Platonova, Elena; Kobylkov, Dmitry; Vakoliuk, Irina; Valkiūnas, Gediminas
Avian malaria parasites (Haemosporida, Plasmodium) are of cosmopolitan distribution, and they have a significant impact on vertebrate host fitness. Experimental studies show that high parasitemia often develops during primary malaria infections. However, field studies only occasionally reveal high parasitemia in free-living birds sampled using the traditional methods of mist-netting or trapping, and light chronic infections predominate. The reason for this discrepancy between field observation and experimental data remains insufficiently understood. Since mist-netting is a passive capture method, two main parameters determine its success in sampling infected birds in wildlife, i. e. the presence of parasitized birds at a study site and their mobility. In other words, the trapping probability depends on the survival rate of birds and their locomotor activity during infection. Here we test (1) the mortality rate of wild birds infected with Plasmodium relictum (the lineage pSGS1), (2) the changes in their behaviour during presence of an aerial predator, and (3) the changes in their locomotor activity at the stage of high primary parasitemia.We show that some behavioural features which might affect a bird's survival during a predator attack (time of reaction, speed of flush flight and take off angle) did not change significantly during primary infection. However, the locomotor activity of infected birds was almost halved compared to control (non-infected) birds during the peak of parasitemia. We report (1) the markedly reduced mobility and (2) the 20% mortality rate caused by P. relictum and conclude that these factors are responsible for the underrepresentation of birds in mist nets and traps during the stage of high primary parasitemia in wildlife. This study indicates that the widespread parasite, P. relictum (pSGS1) influences the behaviour of birds during primary parasitemia. Experimental studies combined with field observations are needed to better understand the
Liu, Yuehuan; Yang, Zhiyuan; Wang, Xiuqing; Chen, Jiming; Yao, Jiezhang; Song, Yanjun; Lin, Jian; Han, Chunhua; Duan, Huijuan; Zhao, Jicheng; Pan, Jie; Xie, Jia
To determine the susceptibility of pigeons to the newly emerged avian influenza virus subtype H7N9, we experimentally infected three different types of pigeons (meat, town, and racing) with two different doses (2 × 10(4) or 2 × 10(5) EID50) of H7N9 avian influenza virus A/Chicken/China/2013 by either intranasal and intraocular inoculation (IN + IO) or intravenous injection (IV). In addition, the potential transmission of H7N9 to pigeons by direct close contact with experimentally infected pigeons and chickens was assessed. Results showed that none of the experimentally infected pigeons exhibited any clinical signs regardless of the infection route and dose. Of the 12 racing pigeons that were randomly selected and necropsied, none of them had any gross lesions. In agreement with this finding, virus was not isolated from all pigeons. No detectable H7-specific antibodies were found in any pigeon. In contrast, 11 of 31 chickens that were either directly infected with H7N9 by IN + IO inoculation or by contact with IN + IO-infected chickens had conjunctivitis. Virus was isolated from all 31 chickens and H7-specific antibodies were detected in these chickens. However, none of the IV-infected chickens or chickens in direct contact with IV-infected chickens had any clinical signs. No virus was isolated from these chickens and no H7-specific antibody was detected. Overall, we conclude that pigeons are less or not susceptible to the H7N9 virus at the doses used and are not likely to serve as a reservoir for the virus. However, the virus does cause conjunctivitis in chickens and can transmit to susceptible hosts by direct contact.
Peters, Iain R.; Helps, Chris R.; Willi, Barbara; Hofmann-Lehmann, Regina; Gruffydd-Jones, Timothy J.; Day, Michael J.; Tasker, Séverine
The aim of this study was to use fluorescence in-situ hybridisation (FISH) to search for the tissues and cell types important in survival and persistence of Mycoplasma haemofelis, “Candidatus Mycoplasma haemominutum” or “Candidatus Mycoplasma turicensis” in infected cats. A 16S rDNA probe for each species was applied to formalin-fixed, paraffin wax-embedded tissues sections collected from experimentally infected cats. Tissues (n = 12) were collected, at necropsy, from ten cats which had been infected with M. haemofelis, and one each with “Ca. M. haemominutum” and “Ca. M. turicensis”. M. haemofelis specific hybridisation was present on red blood cells (RBCs) in all tissues from acutely infected cats, but not the majority of tissues from chronically infected cats. “Ca. M. haemominutum” specific hybridisation was present on scattered RBCs within the spleen and liver. Specific probe hybridisation was not detected in any of the “Ca. M. turicensis” infected tissues. Haemoplasmas were detected on the surface of RBCs only and not any other cell type. Additionally, FISH was limited by sensitivity and could not detect the lower numbers of organisms present in tissues of cats chronically infected with M. haemofelis. Occasional organisms were detected in cats acutely infected with “Ca. M. haemominutum” but not “Ca. M. turicensis”. PMID:21129480
Yorinks, N.; Atkinson, C.T.
We used behavioral, physiological, and parasitological measures to document effects of acute malarial infections on activity budgets of experimentally infected juvenile Apapane (Himatione sanguinea). Five of eight birds died within 20 to 32 days after exposure to a single infective mosquito bite. Infected Apapane devoted less time to locomotory activities involving flight, walking or hopping, and stationary activities such as singing, preening, feeding, and probing. The amount of time spent sitting was positively correlated with parasitemia and increased dramatically after infection and between treatment and control groups. Birds that succumbed to infection experienced a significant loss of body mass and subcutaneous fat, whereas surviving Apapane were better able to maintain body condition and fat levels. When rechallenged with the parasite five months after initial infection, surviving birds experienced no increase in parasitemia, indicating that they had become immune to reinfection. Regardless of the outcome, infected birds experienced acute illness that would have left them unable to forage or to escape from predators in the wild.
Zheng, F Y; Chen, Q W; Li, Z; Gong, X W; Wang, J D; Yin, H
Bovine ephemeral fever (BEF) is an arthropod-borne viral disease that occurs throughout mainland China. LS11 obtained in the 2011 BEF epidemic was a wild strain, and its virulence and antibody response have never been studied in China. Therefore, the issues were investigated in this work. Experimental cattle were intravenously infected with different doses of BEF virus, and some non-infected cattle were simultaneously monitored. Blood and serum samples were collected from all animals over the course of our study. Infected cattle were challenged for a second time with BEF virus to determine protective period of the antibodies. BEF virus was detected in blood samples from infected cattle, but not in monitored cattle. The neutralizing antibodies (nAbs) against BEFV were easier to be detected and persisted for longer periods in cattle infected with higher doses of BEFV than in those infected with lower doses. When the titer of nAbs was equal to 5 or 6, re-infected cattle still could mount a challenge against BEFV. However, after 3 or 6months, when nAbs were no longer apparent, re-infected cattle displayed typical symptoms of BEF. Our findings indicated that vaccination should be performed once the titer of nAb decreased to 5 or 6.
Magombedze, Gesham; Shiri, Tinevimbo; Eda, Shigetoshi; Stabel, Judy R.
Available diagnostic assays for Mycobacterium avium subsp. paratuberculosis (MAP) have poor sensitivities and cannot detect early stages of infection, therefore, there is need to find new diagnostic markers for early infection detection and disease stages. We analyzed longitudinal IFN-γ, ELISA-antibody and fecal shedding experimental sensitivity scores for MAP infection detection and disease progression. We used both statistical methods and dynamic mathematical models to (i) evaluate the empirical assays (ii) infer and explain biological mechanisms that affect the time evolution of the biomarkers, and (iii) predict disease stages of 57 animals that were naturally infected with MAP. This analysis confirms that the fecal test is the best marker for disease progression and illustrates that Th1/Th2 (IFN-γ/ELISA antibodies) assays are important for infection detection, but cannot reliably predict persistent infections. Our results show that the theoretical simulated macrophage-based assay is a potential good diagnostic marker for MAP persistent infections and predictor of disease specific stages. We therefore recommend specifically designed experiments to test the use of a based assay in the diagnosis of MAP infections. PMID:28317944
Horta, Aline Luciano; Leite, Ana Luisa Junqueira; Paula Costa, G.
Trypanosoma cruzi causes a cardiac infection characterized by an inflammatory imbalance that could become the inciting factor of the illness. To this end, we evaluated the role of carvedilol, a beta-blocker with potential immunomodulatory properties, on the immune response in C57BL/6 mice infected with VL-10 strain of T. cruzi in the acute phase. Animals (n = 40) were grouped: (i) not infected, (ii) infected, (iii) infected + carvedilol, and (iv) not infected + carvedilol. We analyzed parameters related to parasitemia, plasma levels of TNF, IL-10, and CCL2, and cardiac histopathology after the administration of carvedilol for 30 days. We did not observe differences in the maximum peaks of parasitemia in the day of their detection among the groups. The plasma TNF was elevated at 60 days of infection in mice treated or not with carvedilol. However, we observed a decreased CCL2 level and increased IL-10 levels in those infected animals treated with carvedilol, which impacted the reduction of the inflammatory infiltration in cardiac tissue. For this experimental model, carvedilol therapy was not able to alter the levels of circulating parasites but modulates the pattern of CCL2 and IL-10 mediators when the VL10 strain of T. cruzi was used in C57BL6 mice. PMID:28377930
Magombedze, Gesham; Shiri, Tinevimbo; Eda, Shigetoshi; Stabel, Judy R.
Available diagnostic assays for Mycobacterium avium subsp. paratuberculosis (MAP) have poor sensitivities and cannot detect early stages of infection, therefore, there is need to find new diagnostic markers for early infection detection and disease stages. We analyzed longitudinal IFN-γ, ELISA-antibody and fecal shedding experimental sensitivity scores for MAP infection detection and disease progression. We used both statistical methods and dynamic mathematical models to (i) evaluate the empirical assays (ii) infer and explain biological mechanisms that affect the time evolution of the biomarkers, and (iii) predict disease stages of 57 animals that were naturally infected with MAP. This analysis confirms that the fecal test is the best marker for disease progression and illustrates that Th1/Th2 (IFN-γ/ELISA antibodies) assays are important for infection detection, but cannot reliably predict persistent infections. Our results show that the theoretical simulated macrophage-based assay is a potential good diagnostic marker for MAP persistent infections and predictor of disease specific stages. We therefore recommend specifically designed experiments to test the use of a based assay in the diagnosis of MAP infections.
Sarrazin, S; Dewulf, J; Mathijs, E; Laureyns, J; Mostin, L; Cay, A B
Bovine viral diarrhoea virus (BVDV) causes persistent infections by infecting the fetus of susceptible animals during gestation. These persistently infected (PI) animals are important sources of infection. On the contrary, transiently infected (TI) animals are believed to be less important, but transient infections with a severe BVDV-2 strain can spread explosively. To assess the importance of TI cattle in the epidemiology of BVDV, two experimental infections were performed to determine basic reproduction ratios (R0). In each experiment three calves were infected via intranasal inoculation and housed together with seven susceptible animals. Two strains isolated in Belgium were used, a virulent BVDV-1b and a virulent BVDV-2a field isolate, resulting in an R0 of 0.25 (95% CI 0.01; 1.95) and 0.24 (95% CI 0.01; 2.11), respectively. A PI animal was then introduced to the remaining uninfected animals and produced an R of +∞ (95% CI 1.88; +∞). These results support the suggestion that TI animals, compared to PI animals, contribute only a limited amount to BVDV spread. Additionally, the severe clinical symptoms observed in the field with these isolates could not be reproduced during these experiments, suggesting that other factors besides strain virulence influence the clinical manifestations evoked by BVDV.
Witonsky, Sharon G; Ellison, Siobhan; Yang, Jibing; Gogal, Robert M; Lawler, Heather; Suzuki, Yasuhiro; Sriranganathan, Namalwar; Andrews, Frank; Ward, Daniel; Lindsay, David S
Equine protozoal myeloencephalitis (EPM) due to Sarcocystis neurona infection is 1 of the most common neurologic diseases in horses in the United States. The mechanisms by which most horses resist disease, as well as the possible mechanisms by which the immune system may be suppressed in horses that develop EPM, are not known. Therefore, the objectives of this study were to determine whether horses experimentally infected with S. neurona developed suppressed immune responses. Thirteen horses that were negative for S. neurona antibodies in serum and cerebrospinal fluid (CSF) were randomly assigned to control (n = 5) or infected (n = 8) treatment groups. Neurologic exams and cerebrospinal fluid analyses were performed prior to, and following, S. neurona infection. Prior to, and at multiple time points following infection, immune parameters were determined. All 8 S. neurona-infected horses developed clinical signs consistent with EPM, and had S. neurona antibodies in the serum and CSF. Both infected and control horses had increased percentages (P < 0.05) of B cells at 28 days postinfection. Infected horses had significantly decreased (P < 0.05) proliferation responses as measured by thymidine incorporation to nonspecific mitogens phorbol myristate acetate (PMA) and ionomycin (I) as soon as 2 days postinfection.
Hernández-Jáuregui, P; Ramírez Mendoza, H; Mercado García, C; Moreno-López, J; Kennedy, S
Porcine rubulavirus (La Piedad-Michoacan virus) (PoRV-LPMV) is a member of the Paramyxoviridae family that causes encephalitis in young piglets and infertility in adult sows and boars. Infertility in sows naturally infected by PoRV-LPMV is characterized by an increased number of returns to oestrus, stillbirths and mummified fetuses. In this study, nine seronegative gilts were inoculated intranasally with the PAC-3 strain of PoRV-LPMV at week 6 or 10 of gestation. These animals were then killed at weeks 8 or 15 of gestation (seven gilts) or after natural parturition (two gilts). Four control gilts were mock-infected at gestation week 6 or 10 and killed between 2 and 4 weeks later. Gross lesions of focal congestion and haemorrhage were seen in the placenta and endometrium of one gilt infected at gestation week 6 and one infected at gestation week 10. PoRV-LPMV was isolated, at 2-6 weeks post-inoculation (pi), from lung, tonsils, ovary, placenta, uterus and lymph nodes of three of the gilts infected at gestation week 6 and at 2-3 weeks pi from lung, tonsil and ovary of two gilts infected at gestation week 10. Many of the fetuses of eight infected gilts were smaller than normal and had dermal ecchymoses. Dehydrated or mummified fetuses were present in six of the infected gilts but not in any control animal. PoRV-LPMV was isolated from brain, lung and liver of fetuses from two gilts infected at gestation week 6, and from two infected at gestation week 10. These results indicate that, after experimental infection, PoRV can replicate in tissues of seronegative pregnant gilts, cross the placenta, and cause fetal death and mummification.
Neul, Annkatrin; Schrödl, Wieland; Marschang, Rachel E; Bjick, Tina; Truyen, Uwe; von Buttlar, Heiner; Pees, Michael
OBJECTIVE To measure immunologic responses of snakes after experimentally induced infection with ferlaviruses. ANIMALS 42 adult corn snakes (Pantherophis guttatus) of both sexes. PROCEDURES Snakes were inoculated intratracheally with genogroup A (n = 12), B (12), or C (12) ferlavirus (infected groups) or cell-culture supernatant (6; control group) on day 0. Three snakes from each infected group were euthanized on days 4, 16, 28, and 49, and 3 snakes from the control group were euthanized on day 49. Blood samples were collected from live snakes on days -6 (baseline), 4, 16, 28, and 49. Hematologic tests were performed and humoral responses assessed via hemagglutination-inhibition assays and ELISAs. Following euthanasia, gross pathological and histologic evaluations and virus detection were performed. RESULTS Severity of clinical signs of and immunologic responses to ferlavirus infection differed among snake groups. Hematologic values, particularly WBC and monocyte counts, increased between days 4 and 16 after infection. A humoral response was identified between days 16 and 28. Serum IgM concentrations increased from baseline earlier than IgY concentrations, but the IgY relative increase was higher at the end of the study. The hemagglutination-inhibition assay revealed that the strongest reactions in all infected groups were against the strain with which they had been infected. Snakes infected with genogroup A ferlavirus had the strongest immune response, whereas those infected with genogroup B had the weakest responses. CONCLUSIONS AND CLINICAL RELEVANCE Results of this experimental study suggested that the ferlavirus strain with the highest virulence induced the weakest immune response in snakes.
Jaenichen, Hans-Rainer; Pitz, Johann
In the public debate about patents, specifically in the area of biotechnology, the position has been taken that patents block the progress of science. As we demonstrate in this review, this is not the case in the European Union (EU). The national patent acts of the EU member states define research and experimental use exemptions from patent infringement that allow sufficient room for research activities to promote innovation. This review provides a comparative overview of the legal requirements and the extent and limitations of experimental use exemptions, including the so-called Bolar provision, in Germany, the United Kingdom, France, Spain, Italy, and The Netherlands. The legal framework in the respective countries is illustrated with reference to practical examples concerning tests on patent-protected genetic targets and antibodies. Specific questions concerning the use of patent-protected research tools, the outsourcing of research activities, and the use of preparatory and supplying acts for experimental purposes that are necessary for conducting experiments are covered. PMID:25377145
Jaenichen, Hans-Rainer; Pitz, Johann
In the public debate about patents, specifically in the area of biotechnology, the position has been taken that patents block the progress of science. As we demonstrate in this review, this is not the case in the European Union (EU). The national patent acts of the EU member states define research and experimental use exemptions from patent infringement that allow sufficient room for research activities to promote innovation. This review provides a comparative overview of the legal requirements and the extent and limitations of experimental use exemptions, including the so-called Bolar provision, in Germany, the United Kingdom, France, Spain, Italy, and The Netherlands. The legal framework in the respective countries is illustrated with reference to practical examples concerning tests on patent-protected genetic targets and antibodies. Specific questions concerning the use of patent-protected research tools, the outsourcing of research activities, and the use of preparatory and supplying acts for experimental purposes that are necessary for conducting experiments are covered.
Valerio, Lluís; Vallès, Xavier; Morales, Betty; Garcia-Diaz, M Immaculada; Pedro-Botet, M Luisa; Serra, Jordi
Background Immigration-related new diseases pose a growing challenge for healthcare services in receptor countries. Following Latin American migration, Chagas disease has inevitably appeared in Europe. Aim To determine the prevalence and characteristics of oesophageal motility disorders in immigrants infected with Trypanosoma cruzi, using high resolution oesophageal manometry (HREM). Methods In all newly-diagnosed cases with chronic Chagas infection referring upper digestive symptoms, a protocolized clinical evaluation and complementary tests including barium oesophagogram and HREM were carried out. As control group, 14 healthy subjects from the same endemic areas were studied with HREM. Results We included 61 patients (46 female, 15 male; age range 26–63 years). Only seven patients (11%) had a minor alteration on barium oesophagogram. By contrast, 23 (37%) patients showed an alteration in oesophageal manometry, mainly minor motility disorders (34%). Only one healthy control (7%) had a minor motility disorder at HREM (p = 0.029 vs. patients). Conclusions Oesophageal motor disorders in infected immigrants with Chagas disease are common, and mainly characterized by a minor motility disorder that is not detected by barium oesophagogram. Hence, as well as barium oesophagogram examination, HREM should be considered, to assess oesophageal damage in this specific group of patients. PMID:27536373
von Bernuth, Horst; Picard, Capucine; Puel, Anne; Casanova, Jean-Laurent
Most Toll-like-receptors (TLRs) and interleukin-1 receptors (IL-1Rs) signal via myeloid differentiation primary response 88 (MyD88) and interleukin-1 receptor-associated kinase 4 (IRAK-4). The combined roles of these two receptor families in the course of experimental infections have been assessed in MyD88- and IRAK-4-deficient mice for almost fifteen years. These animals have been shown to be susceptible to 46 pathogens: 27 bacteria, 8 viruses, 7 parasites, and 4 fungi. Humans with inborn MyD88 or IRAK-4 deficiency were first identified in 2003. They suffer from naturally occurring life-threatening infections caused by a small number of bacterial species, although the incidence and severity of these infections decrease with age. Mouse TLR- and IL-1R-dependent immunity mediated by MyD88 and IRAK-4 seems to be vital to combat a wide array of experimentally administered pathogens at most ages. By contrast, human TLR- and IL-1R-dependent immunity mediated by MyD88 and IRAK-4 seems to be effective in the natural setting against only a few bacteria and is most important in infancy and early childhood. The roles of TLRs and IL-1Rs in protective immunity deduced from studies in mutant mice subjected to experimental infections should therefore be reconsidered in the light of findings for natural infections in humans carrying mutations as discussed in this review. PMID:23255009
Kuhn, Thomas; Benninghoff, Tim; Karl, Horst; Landry, Thomas; Klimpel, Sven
European smelt Osmerus eperlanus (n = 501) from the German Wadden Sea (North Sea) near the city of Cuxhaven were examined for their infestation with parasitic anisakid nematodes, especially with sealworms of the genus Pseudoterranova. The distribution of third-stage larvae (L3) in the musculature and viscera of the fish was analyzed. In total, we isolated 543 L3 from the hosts' body cavity and musculature. A subsample of 105 larvae were identified as the (sibling) species P. decipiens s.s. using direct sequencing of the highly variable ribosomal ITS1-5.8S-ITS2 genetic marker. The mean abundance was 1.1, the mean intensity was 2.3 P. decipiens s.s. and the prevalence was 47.3%. Total length and total weight, but not Fulton's condition factor (K), were significantly different in infected compared to uninfected smelt. No correlation was found between the total length of infected fish and the intensity of anisakid nematodes. The vast majority of P. decipiens s.s. was found in the musculature of the smelt. More than half (55.7%) of all nematodes were located in the 3 parts of the epaxial musculature, whereas 18.4 and 26.0% were found in the hypaxial musculature and the compartments of the tail muscles, respectively.
Tenke, Peter; Kovacs, Bela; Bjerklund Johansen, Truls E; Matsumoto, Tetsuro; Tambyah, Paul A; Naber, Kurt G
We surveyed the extensive literature regarding the development, therapy and prevention of catheter-associated urinary tract infections (UTIs). We systematically searched for meta-analyses of randomised controlled trials available in Medline giving preference to the Cochrane Central Register of Controlled Trials and also considered other relevant publications, rating them on the basis of their quality. The studies' recommendations, rated according to a modification of the US Department of Health and Human Services (1992), give a close-to-evidence-based guideline for all medical disciplines, with special emphasis on urology where catheter care is an important issue. The survey found that the urinary tract is the commonest source of nosocomial infection, particularly when the bladder is catheterised (IIa). Most catheter-associated UTIs are derived from the patient's own colonic flora (IIb) and the catheter predisposes to UTI in several ways. The most important risk factor for the development of catheter-associated bacteriuria is the duration of catheterisation (IIa). Most episodes of short-term catheter-associated bacteriuria are asymptomatic and are caused by a single organism (IIa). Further organisms tend to be acquired by patients catheterised for more than 30 days. The clinician should be aware of two priorities: the catheter system should remain closed and the duration of catheterisation should be minimal (A). While the catheter is in place, systemic antimicrobial treatment of asymptomatic catheter-associated bacteriuria is not recommended (A), except for some special cases. Routine urine culture in an asymptomatic catheterised patient is also not recommended (C) because treatment is in general not necessary. Antibiotic treatment is recommended only for symptomatic infection (B). Long-term antibiotic suppressive therapy is not effective (A). Antibiotic irrigation of the catheter and bladder is of no advantage (A). Routine urine cultures are not recommended if the
Penzhorn, B L; Lewis, B D; López-Rebollar, L M; Swan, G E
The efficacy of 5 drugs was tested against experimental Babesia felis infection in domestic cats. Two of the drugs, rifampicin and a sulphadiazine-trimethoprim combination, appeared to have an anti-parasitic effect, but were inferior to primaquine. The other 3 drugs, buparvaquone, enrofloxacin and danofloxacin, had no significant anti-babesial effect.
The current study was designed to compare experimental oral and intraperitoneal inoculation on early host immune responses to MAP infection. Blood samples were obtained on d -5 and -4, 7, 14, 21, 28, and monthly thereafter for the 12 month term of the study. Isolation of peripheral blood mononuclear...
Understanding the infection process to Mycobacterium avium subsp. paratuberculosis is tantamount to the development of effective vaccines and therapeutics for the control of this disease in the field. The current study compared the effectiveness of oral and intraperitoneal methods of experimental in...
Effect of Sericea lespedeza (SL; Lespedeza cuneata) leaf meal feeding was evaluated in two experiments in indoor reared goats with experimental infection of Haemonchus contortus (HC) larvae. In the first experiment, ten, 8-10 months old male kids from Spanish and Alpine cross bred, pair matched for...
Tevebaugh, M. D.; Nelson, J. P.
The effect is reported to design, fabricate, test and evaluate a prototype experimental system for the control of surgically induced infections. The purpose is to provide the cleanest possible environment within a hospital surgery room and eliminate contamination sources that could cause infections during surgery. The system design is described. The system provides for a portable laminar flow clean room, a full bubble helmet system with associated communications and ventilation subsystems for operating room personnel, and surgical gowns that minimize the migration of bacteria. The development test results consisting of portability, laminar flowrate, air flow pattern, electrostatic buildup, noise level, ventilation, human factors, electrical and material compatibility tests are summarized. The conclusions are that the experimental system is effective in reducing the airborne and wound contamination although the helmets and gowns may not be a significant part of this reduction. Definitive conclusions with regard to the infection rate cannot be made at this time.
Engelsman, Anton F; Saldarriaga-Fernandez, Isabel C; Nejadnik, M Reza; van Dam, Gooitzen M; Francis, Kevin P; Ploeg, Rutger J; Busscher, Henk J; van der Mei, Henny C
The fate of secondary biomaterial implants was determined by bio-optical imaging and plate counting, after antibiotic treatment of biomaterials-associated-infection (BAI) and surgical removal of an experimentally infected, primary implant. All primary implants and tissue samples from control mice showed bioluminescence and were culture-positive. In an antibiotic treated group, no bioluminescence was detected and only 20% of all primary implants and no tissue samples were culture-positive. After revision surgery, bioluminescence was detected in all control mice. All the implants and 80% of all tissue samples were culture-positive. In contrast, in the antibiotic treated group, 17% of all secondary implants and 33% of all tissue samples were culture-positive, despite antibiotic treatment. The study illustrates that due to the BAI of a primary implant, the infection risk of biomaterial implants is higher in revision surgery than in primary surgery, emphasizing the need for full clearance of the infection, as well as from surrounding tissues prior to implantation of a secondary implant.
Coelho, Ana Maria M.; Sampietre, Sandra; Patzina, Rosely; Jukemura, Jose; Cunha, Jose Eduardo M.; Machado, Marcel C.C.
Objective. Acute pancreatitis is one the important causes of systemic inflammatory response syndrome (SIRS). SIRS results in gut barrier dysfunction that allows bacterial translocation and pancreatic infection to occur. Indomethacin has been used to reduce inflammatory process and bacterial translocation in experimental models. The purpose of this study was to determine the effect of inhibition of prostaglandin E2 (PGE2) production on pancreatic infection. Materials and methods. An experimental model of severe acute pancreatitis (AP) was utilized. The animals were divided into three groups: sham (surgical procedure without AP induction); pancreatitis (AP induction); and indomethacin (AP induction plus administration of 3 mg/kg of indomethacin). Serum levels of interleukin (IL)-6 and IL-10, PGE2, and tumor necrosis factor (TNF)-α were measured 2 h after the induction of AP. We analyzed the occurrence of pancreatic infection with bacterial cultures performed 24 h after the induction of AP. The occurrence of pancreatic infection (considered positive when the CFU/g was >105), pancreatic histologic analysis, and mortality rate were studied. Results. In spite of the reduction of IL-6, IL-10, and PGE2 levels in the indomethacin group, TNF-α level, bacterial translocation, and pancreatic infection were not influenced by administration of indomethacin. The inhibition of PGE2 production did not reduce pancreatic infection, histologic score, or mortality rate. Conclusion. The inhibition of PGE2 production was not able to reduce the occurrence of pancreatic infection and does not have any beneficial effect in this experimental model. Further investigations will be necessary to discover a specific inhibitor that would make it possible to develop an anti-inflammatory therapy. PMID:18345325
Kim, Woo Sik; Kim, Jong-Seok; Kim, Hong Min; Kwon, Kee Woong; Cho, Sang-Nae; Shin, Sung Jae; Koh, Won-Jung
The latency and reactivation of Mycobacterium tuberculosis infection has been well studied. However, there have been few studies of the latency and reactivation of Mycobacterium avium complex (MAC), the most common etiological non-tuberculous Mycobacterium species next to M. tuberculosis in humans worldwide. We hypothesized that latent MAC infections can be reactivated following immunosuppression after combination chemotherapy with clarithromycin and rifampicin under experimental conditions. To this end, we employed a modified Cornell-like murine model of tuberculosis and investigated six strains consisting of two type strains and four clinical isolates of M. avium and M. intracellulare. After aerosol infection of each MAC strain, five to six mice per group were euthanized at 2, 4, 10, 18, 28 and 35 weeks post-infection, and lungs were sampled to analyze bacterial burden and histopathology. One strain of each species maintained a culture-negative state for 10 weeks after completion of 6 weeks of chemotherapy, but was reactivated after 5 weeks of immunosuppression in the lungs with dexamethasone (three out of six mice in M. avium infection) or sulfasalazine (four out of six mice in both M. avium and M. intracellulare infection). The four remaining MAC strains exhibited decreased bacterial loads in response to chemotherapy; however, they remained at detectable levels and underwent regrowth after immunosuppression. In addition, the exacerbated lung pathology demonstrated a correlation with bacterial burden after reactivation. In conclusion, our results suggest the possibility of MAC reactivation in an experimental mouse model, and experimentally demonstrate that a compromised immune status can induce reactivation and/or regrowth of MAC infection. PMID:26406237
Rolán, Hortensia G; den Hartigh, Andreas B; Kahl-McDonagh, Melissa; Ficht, Thomas; Adams, L Garry; Tsolis, Renée M
The Brucella species type IV secretion system, encoded by the virB1-12 locus, is required for intracellular replication and persistent infection in vivo. The requirement of VirB proteins for infection suggests that they are expressed in vivo and may therefore represent serological markers of infection. To test this idea, we purified recombinant VirB1, VirB5, VirB11, and VirB12 and tested for their recognition by antibodies in sera from experimentally infected mice and goats by using an indirect enzyme-linked immunosorbent assay. Antibody responses to VirB12 but not to VirB1, VirB5, or VirB11 were detected in 20/20 mice experimentally inoculated with Brucella abortus and 12/12 goats experimentally infected with Brucella melitensis. The potential use of VirB12 as a serological tool for the diagnosis of brucellosis was evaluated in the natural bovine host. Serum samples from 145 cattle of known serology (29% negative and 71% positive) were analyzed for the production of antibody responses to VirB12. One hundred two cattle samples (70.3%) were positive for antibodies to VirB12, while 43 samples were negative (29.7%). A positive serological response to VirB12 correlated with positive serology to whole B. abortus antigen in 99% of samples tested. These results show that VirB12 is expressed during infection of both experimental and natural hosts of Brucella species, and they suggest that VirB12 may be a useful serodiagnostic marker for brucellosis.
Amman, Brian R; Jones, Megan E B; Sealy, Tara K; Uebelhoer, Luke S; Schuh, Amy J; Bird, Brian H; Coleman-McCray, JoAnn D; Martin, Brock E; Nichol, Stuart T; Towner, Jonathan S
Marburg virus (Marburg marburgvirus; MARV) causes sporadic outbreaks of Marburg hemorrhagic fever (MHF) in Africa. The Egyptian fruit bat (Rousettus aegyptiacus) has been identified as a natural reservoir based most-recently on the repeated isolation of MARV directly from bats caught at two locations in southwestern Uganda where miners and tourists separately contracted MHF from 2007-08. Despite learning much about the ecology of MARV through extensive field investigations, there remained unanswered questions such as determining the primary routes of virus shedding and the severity of disease, if any, caused by MARV in infected bats. To answer these questions and others, we experimentally infected captive-bred R. aegyptiacus with MARV under high (biosafety level 4) containment. These experiments have shown infection profiles consistent with R. aegyptiacus being a bona fide natural reservoir host for MARV and demonstrated routes of viral shedding capable of infecting humans and other animals.
Materniak, Magdalena; Hechler, Torsten; Löchelt, Martin; Kuzmak, Jacek
Foamy viruses (FVs) are the least known retroviruses commonly found in primates, cats, horses, and cattle. Although FVs are considered apathogenic, simian and feline FVs have been shown to be associated with some transient health abnormalities in animal models. Currently, data regarding the course of infection with bovine FV (BFV) are not available. In this study, we conducted experimental infections of natural (cattle) and heterologous (sheep) hosts with the BFV(100) isolate and monitored infection patterns in both hosts during the early phase postinoculation as well as after long-term infection. Four calves and six sheep inoculated with BFV(100) showed no signs of pathology but developed persistent infection, as confirmed by virus rescue, consistent detection of BFV-specific antibodies, and presence of viral DNA. In both hosts, antibodies against BFV Gag and Bet appeared early after infection and persisted at high and stable levels while seroreactivity toward Env was consistently detectable only in BFV-infected sheep. Interestingly, the BFV proviral DNA load was highest in lung, spleen, and liver and moderate in leukocytes, while salivary glands contained either low or undetectable DNA loads in calves or sheep, respectively. Additionally, comparison of partial BFV sequences from inoculum and infected animals demonstrated very limited changes after long-term infection in the heterologous host, clearly less than those found in BFV field isolates. The persistence of BFV infection in both hosts suggests full replication competence of the BFV(100) isolate with no requirement of genetic adaptation for productive replication in the authentic and even in a heterologous host.
Skouras, Andreas; Schmidt, Verena; Körting, Wolfgang; Steinhagen, Dieter
In a field study, infecting European flounder (Platichthys flesus L.) subclinically with different parasite species did not result in any alteration of the innate immune response. Due to the high variability in infection status and the immune parameters measured, no relationships of biological significance were found. The data indicate that copepods, as the most abundant parasites, most probably had no major influence on immune responses measured here. Thus it might be concluded that these parameters were not sensitive to parasite infections occurring under natural conditions. The immune parameters considered here are regarded as promising indicators of chemical contaminant-induced variation in piscine immune responses, which could be implemented in pollution monitoring programmes.
LESCANO, Susana A. Zevallos; dos SANTOS, Sergio Vieira; ASSIS, Jesiel Maurício Lemos; CHIEFFI, Pedro Paulo
SUMMARY The efficacy of nitazoxanide (NTZ) against toxocariasis was investigated in an experimental murine model and results were compared to those obtained using mebendazole. Sixty male BALB/c mice, aged six to eight weeks-old, were divided into groups of 10 each; fifty were orally infected with 300 larvaed eggs of T. canisand grouped as follows, G I: infected untreated mice; G II: infected mice treated with MBZ (15 mg/kg/day) 10 days postinfection (dpi); G III: infected mice treated with NTZ (20 mg/kg/day) 10 dpi; G IV: infected mice treated with MBZ 60 dpi; G V: infected mice treated with NTZ 60 dpi; GVI: control group comprising uninfected mice. Mice were bled via retro-orbital plexus on four occasions between 30 and 120 dpi. Sera were processed using the ELISA technique to detect IgG anti- Toxocaraantibodies. At 120 dpi, mice were sacrificed for larval recovery in the CNS, liver, lungs, kidneys, eyes and carcass. Results showed similar levels of anti- ToxocaraIgG antibodies among mice infected but not submitted to treatment and groups treated with MBZ or NTZ, 10 and 60 dpi. Larval recovery showed similar values in groups treated with NTZ and MBZ 10 dpi. MBZ showed better efficacy 60 dpi, with a 72.6% reduction in the parasite load compared with NTZ, which showed only 46.5% reduction. We conclude that administration of these anthelmintics did not modify the humoral response in experimental infection by T. canis. No parasitological cure was observed with either drug; however, a greater reduction in parasite load was achieved following treatment with MBZ. PMID:26422159
Lescano, Susana A Zevallos; Santos, Sergio Vieira dos; Assis, Jesiel Maurício Lemos; Chieffi, Pedro Paulo
The efficacy of nitazoxanide (NTZ) against toxocariasis was investigated in an experimental murine model and results were compared to those obtained using mebendazole. Sixty male BALB/c mice, aged six to eight weeks-old, were divided into groups of 10 each; fifty were orally infected with 300 larvaed eggs of T. canis and grouped as follows, G I: infected untreated mice; G II: infected mice treated with MBZ (15 mg/kg/day) 10 days postinfection (dpi); G III: infected mice treated with NTZ (20 mg/kg/day) 10 dpi; G IV: infected mice treated with MBZ 60 dpi; G V: infected mice treated with NTZ 60 dpi; GVI: control group comprising uninfected mice. Mice were bled via retro-orbital plexus on four occasions between 30 and 120 dpi. Sera were processed using the ELISA technique to detect IgG anti- Toxocara antibodies. At 120 dpi, mice were sacrificed for larval recovery in the CNS, liver, lungs, kidneys, eyes and carcass. Results showed similar levels of anti- Toxocara IgG antibodies among mice infected but not submitted to treatment and groups treated with MBZ or NTZ, 10 and 60 dpi. Larval recovery showed similar values in groups treated with NTZ and MBZ 10 dpi. MBZ showed better efficacy 60 dpi, with a 72.6% reduction in the parasite load compared with NTZ, which showed only 46.5% reduction. We conclude that administration of these anthelmintics did not modify the humoral response in experimental infection by T. canis. No parasitological cure was observed with either drug; however, a greater reduction in parasite load was achieved following treatment with MBZ.
Schennings, Torgny; Farnebo, Filip; Szekely, Laszlo; Flock, Jan-Ingmar
A novel murine experimental wound infection model was used to assess the efficacy of multi-component immunization against Staphylococcus aureus infection. Necrotic lesions were induced in mice with venom from Bothrops asper and infected with a low inoculum, 1 × 10(2) CFU. The wound infection model therefore more resembles a clinical case of S. aureus infection compared with conventional infection models where far more bacteria are required. Before infection, mice were immunized with four recombinant S.aureus proteins expressed from Escherichia coli: (i) domains 1-3 of Extracellular adherence protein (Eap), (ii) Efb - D (fusion protein combining Extracellular fibrinogen binding protein (Efb) and a fibronectin binding domain (D) of the fibronectin binding protein (FnBP) and (iii) clumping factor A (ClfA). In the immunized group, lower bacterial colonization, undisturbed crust formation and significantly faster wound healing were found compared with the unimmunized control group. Efb and Eap have previously been found to impair wound healing and neutralization of these proteins by antibodies restores a more natural wound healing process. This effect is further also enhanced by the proposed opsonic activity of antibodies against ClfA and FnBP.
Veselinova, A; Semov, P
Comparative pathomorphologic studies were carried out with 40 birds (29 turkeys and 11 hens) infected per os with 0.5 cc embryonal fluid 10(5) ELD 50 each with two Newcastle disease virus strains (Texas -- standard and 7/5 isolated from dead hens and typed as viscerotropic). It was found that the pathoanatomical picture is negative for all birds experimentally infected with the Texas strain. Haemorrhagic necrotic modification of the glandular stomach and intestine were found in birds infected with strain 7/5. The pathoanatomical finding is better expressed in hens compared with that for turkeys. The histologic modification in birds infected with strain Texas were manifested mostly by nonsuppurative encephalitis. Haemorrhagic-diphtheroid inflammation of the alimentary tract, haemorrhages in the viscera and unreactive necrotic foci in the spleen as well as poorly manifested inflammatory changes in the central nervous system were found in birds infected with strain 7/5. No pathoanatomical modifications for Newcastle disease were found in turkeys, analogous to hens, infected with velogenic neurotropic virus. The velogenic viscerotropic strain is less pathogenic for turkeys than for hens. It causes slighter haemorrhagic necrotic modification in turkeys viscera than in hens. Nonsuppurative encephalitis established histologically in both turkeys and hens infected with neurotropic and viscerotropic Newcastle disease virus is a constant finding and may be used as a diagnostic sign of the disease.
Luisi, Elena; Forzan, Mario; Giusti, Matteo; Sagona, Simona; Tolari, Francesco; Felicioli, Antonio
Deformed wing virus (DWV) is a honeybee pathogen whose presence is generally associated with infestation of the colony by the mite Varroa destructor, leading to the onset of infections responsible for the collapse of the bee colony. DWV contaminates bee products such as royal jelly, bee-bread and honey stored within the infected hive. Outside the hive, DWV has been found in pollen loads collected directly from infected as well as uninfected forager bees. It has been shown that the introduction of virus-contaminated pollen into a DWV-free hive results in the production of virus-contaminated food, whose role in the development of infected bees from virus-free eggs has been experimentally demonstrated. The aim of this study was twofold: (i) to ascertain the presence of DWV on pollen collected directly from flowers visited by honeybees and then quantify the viral load and (ii) determine whether the virus associated with pollen is infective. The results of our investigation provide evidence that DWV is present on pollen sampled directly from visited flowers and that, following injection in individuals belonging to the pollinator species Apis mellifera, it is able to establish an active infection, as indicated by the presence of replicating virus in the head of the injected bees. We also provide the first indication that the pollinator species Osmia cornuta is susceptible to DWV infection. PMID:25419704
Bangoura, B; Daugschies, A; Fuerll, M
Coccidiosis, often caused by Eimeria zuernii infection, is an important diarrhoeal disease in calves [Fitzgerald, P.R., 1980. The economic impact of coccidiosis in domestic animals. Adv. Vet. Sci. Comp. Med. 24, 121-143]. Infection trials were performed to investigate the effects of experimental E. zuernii coccidiosis on clinical blood chemistry in calves. Three groups of calves were formed: group 1 (n=14) served as uninfected control group, group 2 (n=11) was infected with 150,000 sporulated E. zuernii oocysts per calf, and group 3 (n=16) was infected with 250,000 sporulated E. zuernii oocysts per calf. Measurements throughout the prepatent and the patent period revealed a marked influence of E. zuernii infection on the following parameters: total protein, albumin, urea, bilirubin, creatine kinase, free fatty acid concentration, and cholesterol. Aberrances in these were most pronounced in group 3. No significant and/or distinct changes after infection could be detected in blood glucose concentration. E. zuernii infection impairs intestinal function and induces catabolic metabolism in affected calves. Bilirubin, urea and cholesterol concentration, and creatine kinase activity were particularly affected indicating catabolism of protein and lipids.
Wongrak, Kalyakorn; Gauly, Matthias; Daş, Gürbüz
We investigated whether nematode egg excretion through feces of naturally or experimentally infected chickens follow certain patterns within a day, which may allow determining the most appropriate sampling time for the highest parasite egg concentration. Feces samples (n=864) from chickens (n=36) with naturally occurring mixed nematode infections (trials N1, N2) or with an experimental Ascaridia galli infection (E) were collected quantitatively every 4h for four consecutive days. Number of eggs per gram of feces (EPG) was determined, and accumulative egg output (AEO) at each sampling time as well as total number of eggs excreted within 24h (eggs per day, EPD) were then estimated. At the end of the collection period, the hens were necropsied and their worm burdens determined. Naturally infected hens harbored Heterakis gallinarum (100%), Capillaria spp. (95.7%) and A. galli (91.3%). The experimental A. galli infection produced patent infections in all the birds. In general, both fecal egg concentration (EPG) and the amount of feces increased (P<0.05) sharply from the early morning to early-noon (10:00 a.m.) and remained at a high level until evenings which thereafter decreased to their initial levels during the night both in naturally and experimentally infected birds. This resulted in a more apparent increase or a decrease in AEO at the corresponding time points, respectively, and led to much higher egg excretions during the daytime than the nights. Despite the apparent within day fluctuations in egg excretion, neither EPG (P=0.704) nor AEO (P=0.499) nor EPD (P=0.149) was significantly different among the four collection days. Similarly, there was no significant interaction (P>0.05) between effects of sampling hours and days on EPG and AEO, suggesting the existence of repeatable diurnal fluctuations within each day. Although an association between climatic parameters (e.g., ambient temperature and relative humidity) and the nematode egg excretion was quantified, a
Background Ashworthius sidemi, a blood-sucking nematode, is a primary parasite of Asiatic cervides, primarily sika deer (Cervus nippon). As A. sidemi infections are common in bison, red and roe deer, and gastrointestinal nematodes are often exchanged between animals, it is possible that other farm animals such as cows and sheep that may use the same pastures can be infected. Hence, histopathological changes observed in the walls of the abomasa and duodena of infected wildlife caused by a strong parasite presence may become an important health problem also for farm animals. Methods In the present study, a simple PCR test for the detection of A. sidemi infection in European bison based on DNA from third stage infective larvae (L3) has been optimized. Results The species-specific primers generated a 406 bp fragment, and A. sidemi DNA could be detected at concentrations of 0.1 pg/μl. The specificity of PCR was confirmed by the use of the genomic DNA of adult Ostertagia ostertagi, Haemonchus contortus, Cooperiaoncophora as negative controls. Conclusion It is possible to detect A. sidemi infection in European bison using DNA from L3. If this nematode infection is transmitted to cows this method may be effective to diagnose invasion in breeding animals in vivo. PMID:24886355
Galland, Claire; Dupuy, Célie; Loizeau, Véronique; Danion, Morgane; Auffret, Michel; Quiniou, Louis; Laroche, Jean; Pichereau, Vianney
Platichthys flesus is often used as a sentinel species to monitor the estuarine water quality. In this study, we carried out an experimental contamination of fish using a PAHs/PCBs mixture, which was designed to mimic the concentrations found in the Seine estuary (C1) and 10 times these concentrations (C2). We used a proteomic approach to understand the molecular mechanisms implied in the response of P. flesus to these xenobiotics. We showed that 54 proteins were differentially accumulated in one or several conditions, which 34 displayed accumulation factors higher than two. 18 of these proteins were identified by MALDI TOF-TOF mass spectrometry. The results indicated the deregulation of oxidative stress- and glutathione metabolism-(GST, GPx) proteins as well as of several proteins belonging to the betaine demethylation pathway and the methionine cycle (BHMT, SHMT, SAHH), suggesting a role for these different pathways in the P. flesus response to chemical contamination.
Segarra, Amélie; Faury, Nicole; Pépin, Jean-François; Renault, Tristan
Massive mortality outbreaks have been reported in France since 2008 among Pacific oysters, Crassostrea gigas, with the detection of a particular OsHV-1 variant called μVar. Virus infection can be induced in healthy spat in experimental conditions allowing to better understand the disease process, including viral gene expression. Although gene expression of other herpesviruses has been widely studied, we provide the first study following viral gene expression of OsHV-1 over time. In this context, an in vivo transcriptomic study targeting 39 OsHV-1 genes was carried out during an experimental infection of Pacific oyster spat. For the first time, several OsHV-1 mRNAs were detected by real-time PCR at 0 h, 2 h, 4 h, 18 h, 26 h and 42 h post-injection. Several transcripts were detected at 2h post-infection and at 18 h post-infection for all selected ORFs. Quantification of virus gene expression at different times of infection was also carried out using an oyster housekeeping gene, Elongation factor. Developing an OsHV-1-specific reverse transcriptase real time PCR targeting 39 viral gene appears a new tool in terms of diagnosis and can be used to complement viral DNA detection in order to monitor viral replication.
Rieger, J; Janczyk, P; Hünigen, H; Plendl, J
Salmonella Typhimurium is one of the main pathogens compromising porcine and human health as well as food safety, because it is a prevailing source of foodborne infections due to contaminated pork. A prominent problem in the management of this bacteriosis is the number of subclinically infected carrier pigs. As very little is known concerning the mechanisms allowing Salmonella to persist in pigs, the objective of this study was to develop an immunohistochemical approach for the detection of salmonellae in tissue of pigs experimentally infected with Salmonella Typhimurium. Samples were obtained from a challenge trial in which piglets of the German Landrace were intragastrically infected with Salmonella enterica serovar Typhimurium DT104 (1.4-2.1x1010 CFU). Piglets were sacrificed on days 2 and 28 post infection. Tissue samples of jejunum, ileum, colon, ileocecal mesenteric lymph nodes (Lnn. ileocolici), and tonsils (Tonsilla veli palatini) were fixed in Zamboni's fixative and paraffin-embedded. Different immunohistochemical staining protocols were evaluated. Salmonella was detected in varying amounts in the tissues. Brown iron-containing pigments in the lymph nodes interfered with the identification of Salmonella if DAB was used as a staining reagent. Detergents like Triton X-100 or Saponin enhanced the sensitivity. It seems advisable not to use a detection system with brown staining for bacteria in an experimental setup involving intestinal damage including haemorrhage. The use of detergents appears to result in a higher sensitivity in the immunohistochemical detection of salmonellae.
After oral administration of ewes during mid gestation with 2000 freshly prepared sporulated oocysts of T. gondii isolate M4, abortions occurred between days 7 and 11 in 91.6% of pregnant and infected ewes. Afterwards, a further infection was carried out at late gestation in another group of sheep with 500 sporulated oocysts. Abortions happened again between days 9 and 11 post infection (pi) in 58.3% of the infected ewes. Classically, abortions in natural and experimental ovine toxoplasmosis usually occur one month after infection. Few experimental studies have reported the so-called acute phase abortions as early as 7 to 14 days after oral inoculation of oocysts, and pyrexia was proposed to be responsible for abortion, although the underline mechanism was not elucidated. In the present study, all placentas analysed from ewes suffering acute phase abortions showed infarcts and thrombosis in the caruncullar villi of the placentomes and ischemic lesions (periventricular leukomalacia) in the brain of some foetuses. The parasite was identified by PCR in samples from some placentomes of only one sheep, and no antigen was detected by immunohistochemical labelling. These findings suggest that the vascular lesions found in the placenta, and the consequent hypoxic damage to the foetus, could be associated to the occurrence of acute phase abortions. Although the pathogenesis of these lesions remains to be determined, the infectious dose or virulence of the isolate may play a role in their development. PMID:24475786
Hassan, Khaled M; Arafa, Waleed M; Mousa, Waheed M; Shokier, Khaled A M; Shany, Salama A; Aboelhadid, Shawky M
The early detection of Eimeria stiedae in the hepatic tissue of experimentally infected rabbits was investigated using molecular assay. Forty 6-week-old male New Zealand rabbits were divided into two groups. Group A (30 animals) was infected with 2.5 × 10(4) sporulated oocysts of E. stiedae per animal on Day 0 and Group B (10 animals) was used as the uninfected controls. Three animals from Group A and one from Group B were sacrificed at 0, 3, 6, 9, 12, 15, 18, 21, 24 and 27 days post infection (PI). Gross and microscopic post-mortem findings were recorded. Polymerase chain reaction (PCR) of the E. stiedae internal transcribed spacer 1 genomic region was conducted on blood, liver tissue, and feces from the Group A experimentally infected animals. Macroscopically, the liver showed irregular yellowish white nodules pathognomonic to E. stiedae infection beginning on Day 15 PI. Hepatomegaly and ascites were obvious from Day 21-24 PI. The presence of different E. stiedae schizonts and gametocytes in the histopathological sections of the biliary epithelium were evident on Day 15 PI. The E. stiedae PCR was first positive in liver tissues on Day 12 and in fecal samples on Day 18 PI, but the blood samples were negative. In conclusion, the PCR can be used for early diagnosis and control of E. stiedae schizonts before shedding of the oocysts in feces.
Schulz, Claudia; Beer, Martin; Hoffmann, Bernd
During the first epizootic wave of the novel, teratogenic Schmallenberg virus (SBV, Orthobunyavirus) in ruminants in Northern Europe, serological evidence of a previous SBV-infection demonstrated that South American camelids (SAC) are also susceptible to SBV. However, their potential role in SBV spread remains unknown. To investigate the prevalence and course of SBV-infection in SAC, a German field study and an animal trial with three llamas and three alpacas were conducted. From September 2012 to December 2013, 313 of 502 SAC (62.35%) were found SBV seropositive, but negative for SBV-RNA. The estimated between-district (94.23% of 52) and median within-district (71.43%) and herd (73.13%) SBV seroprevalence in German SAC was similar to the seroprevalence reported in cattle herds and sheep flocks at the time. An age of >1 year was found a statistically significant risk factor for SBV-infection, which could be explained by the spatio-temporal spread of SBV in Germany during the study period. No clinical signs or an increase of abortion and congenital malformation associated with SBV-infection in SAC were reported by the study participants. Similar to SBV-infected ruminants, SBV-RNAemia in experimentally SBV-infected SAC was detected for a short time between days 3 and 7 after infection (dpi), and seroconversion occurred between 9 and 21 dpi. Despite the similar virological and serological results, the lack of clinical signs and congenital malformation associated with SBV-infection suggests that SBV causes subclinical infection in SAC. However, their role as reservoirs in the spread of SBV has to be further investigated.
Zhao, Jianjun; Shi, Ning; Sun, Yangang; Martella, Vito; Nikolin, Veljko; Zhu, Chunsheng; Zhang, Hailing; Hu, Bo; Bai, Xue; Yan, Xijun
Canine distemper virus (CDV) infects a broad range of carnivores and causes a highly contagious disease with severe immunosuppression. The disease severity markedly varies in different species. To investigate the pathogenesis of CDV in raccoon dog (Nyctereutes procyonoides), fox (Vulpes vulpes) and mink (Neovison vison) species, three groups of CDV sero-negative animals were infected with CDV strain LN(10)1. This CDV strain belongs to the Asia-1 genotype, which is epidemiologically predominant in carnivores in China. CDV infection provoked marked differences in virulence in the three species that were studied. Raccoon dogs developed fever, severe conjunctivitis, and pathological lesions, with 100% (5/5) mortality and with high viral RNA loads in organs within 15 days post infection (dpi). In infected foxes, the onset of the disease was delayed, with 40% (2/5) mortality by 21 dpi. Infected minks developed only mild clinical signs and pathological lesions, and mortality was not observed. Raccoon dogs and foxes showed more severe immune suppression (lymphopenia, decreased lymphocyte proliferation, viremia and low-level virus neutralizing antibodies) than minks. We also observed a distinct pattern of cytokine mRNA transcripts at different times after infection. Decreased IFN-γ and IL-4 mRNA responses were evident in the animals with fatal disease, while up-regulation of these cytokines was observed in the animals surviving the infection. Increased TNF-α response was detected in animals with mild or severe clinical signs. Based on the results, we could distinguish three different patterns of disease after experimental CDV infection, e.g. a mild form in minks, a moderate form in foxes and a severe disease in raccoon dogs. The observed differences in susceptibility to CDV could be related to distinct host cytokine profiles. Comparative evaluation of CDV pathogenesis in various animal species is pivotal to generate models suitable for the evaluation of CDV
Solórzano, Alicia; Foni, Emanuela; Córdoba, Lorena; Baratelli, Massimiliano; Razzuoli, Elisabetta; Bilato, Dania; Martín del Burgo, María Ángeles; Perlin, David S.; Martínez, Jorge; Martínez-Orellana, Pamela; Fraile, Lorenzo; Chiapponi, Chiara; Amadori, Massimo; del Real, Gustavo
ABSTRACT Avian influenza A viruses have gained increasing attention due to their ability to cross the species barrier and cause severe disease in humans and other mammal species as pigs. H3 and particularly H3N8 viruses, are highly adaptive since they are found in multiple avian and mammal hosts. H3N8 viruses have not been isolated yet from humans; however, a recent report showed that equine influenza A viruses (IAVs) can be isolated from pigs, although an established infection has not been observed thus far in this host. To gain insight into the possibility of H3N8 avian IAVs to cross the species barrier into pigs, in vitro experiments and an experimental infection in pigs with four H3N8 viruses from different origins (equine, canine, avian, and seal) were performed. As a positive control, an H3N2 swine influenza virus A was used. Although equine and canine viruses hardly replicated in the respiratory systems of pigs, avian and seal viruses replicated substantially and caused detectable lesions in inoculated pigs without previous adaptation. Interestingly, antibodies against hemagglutinin could not be detected after infection by hemagglutination inhibition (HAI) test with avian and seal viruses. This phenomenon was observed not only in pigs but also in mice immunized with the same virus strains. Our data indicated that H3N8 IAVs from wild aquatic birds have the potential to cross the species barrier and establish successful infections in pigs that might spread unnoticed using the HAI test as diagnostic tool. IMPORTANCE Although natural infection of humans with an avian H3N8 influenza A virus has not yet been reported, this influenza A virus subtype has already crossed the species barrier. Therefore, we have examined the potential of H3N8 from canine, equine, avian, and seal origin to productively infect pigs. Our results demonstrated that avian and seal viruses replicated substantially and caused detectable lesions in inoculated pigs without previous adaptation
Foreyt, William J; Drew, M L
This experimental study was conducted to evaluate the susceptibility of American bison (Bison bison) to liver flukes, Fascioloides magna and Fasciola hepatica. Six bison were each experimentally inoculated with 600 metacercariae of Fascioloides magna, and three were later treated with triclabendazole suspension at 40 mg/kg of body weight. Four additional bison were each experimentally inoculated with 600 metacercariae of Fasciola hepatica. Five control bison were placebo controls. Two controls and all inoculated bison were euthanized 10 mo (Fascioloides magna) and 7 mo (Fasciola hepatica) after inoculation. None of the control bison or the bison inoculated with Fascioloides magna had flukes or lesions characteristic of fluke infection at necropsy. All four bison inoculated with Fasciola hepatica had characteristic liver fluke lesions at necropsy, and three of four bison contained four, 103, and 111 adult flukes, respectively. Fluke eggs were detected in feces of all Fasciola hepatica-inoculated bison during the experiment, but not from the Fascioloides magna-infected bison or control bison. Clinical signs of infection were not observed during the experiment, but hemoglobin and packed cell volumes were lower in the Fasciola hepatica bison when compared to controls, and eosinophil levels were increased. Triclabendazole at 40 mg/kg of body weight appeared to be safe in bison because no toxic reactions were observed. Results from this study indicated bison are susceptible to infection with Fasciola hepatica and are efficient definitive hosts. Because no Fascioloides magna were recovered, bison may have a decreased susceptibility or innate resistance to Fascioloides magna infection, which may account for a lack of reported infections in this host.
Infection models are useful for studying host responses to infection to aid in the development of diagnostic tools and vaccines. The current study compared experimental oral and intraperitoneal MAP infection on early host immune responses. Twenty neonatal Holstein calves were assigned to 5 treatment...
The objective of this study was to observe early markers of cell-mediated immunity in naïve calves infected with Mycobacterium avium subsp. paratuberculosis (MAP) and how expression of these markers evolved over the 12-month period of infection. Methods of experimental infection included: Control (n...
Zakaryan, Hovakim; Karalova, Elena; Voskanyan, Henrik; Ter-Pogossyan, Zarine; Nersisyan, Narek; Hakobyan, Astghik; Saroyan, David; Karalyan, Zaven
African swine fever is a highly contagious hemorrhagic disease of pigs caused by African swine fever virus (ASFV). Hemorrhages are the most frequently reported lesions in acute and subacute forms of ASF. Hemorrhagic lesions are accompanied by impaired hemostasis, which includes thrombocytopenia and changes in the coagulation system. In the present study, experimental infection was conducted to elucidate whether a highly virulent ASFV genotype II circulating in the Trans-Caucasus and Eastern Europe affects the hemostasis of infected pigs. Platelet count changes and platelet size, as well as coagulation parameters were evaluated upon experimental infection. In contrast to other ASFV strains, ASFV genotype II showed a significant decrease in the number of platelets from 3rd dpi onwards. Furthermore, a decrease in platelet size was observed throughout the entire period of experiment. A significant increase in the number of platelet aggregates was observed from the beginning of infection. Unlike other ASFV strains, ASFV genotype II induced a slight shortening of an activated partial thromboplastin time (aPTT) throughout the experiment. Thrombin time (TT) was prolonged from day 5 onwards, whereas no changes in prothrombin time (PT) were found upon infection. The level of d-dimers was permanently higher than in control with a peak on day 3 post-infection. ASFV induced a significant decrease in the level of fibrinogen from day 5 till the end of experiment. Thus, it can be concluded that ASFV genotype II isolated in Armenia affects the hemostasis of infected pigs and causes changes that differ from that of other ASFV strains described previously.
Batani, D.; Koenig, M.; Baton, S.; Perez, F.; Gizzi, L. A.; Koester, P.; Labate, L.; Honrubia, J.; Debayle, A.; Santos, J.; Schurtz, G.; Hulin, S.; Ribeyre, X.; Fourment, C.; Nicolai, P.; Vauzour, B.; Gremillet, L.; Nazarov, W.; Pasley, J.; Tallents, G.; Richetta, M.; Lancaster, K.; Spindloe, Ch.; Tolley, M.; Neely, D.; Norreys, P.; Kozlova, M.; Nejdl, J.; Rus, B.; Antonelli, L.; Morace, A.; Volpe, L.,; Davies, J.; Wolowski, J.; Badziak, J.
This paper presents the goals and some of the results of experiments conducted within the Working Package 10 (Fusion Experimental Programme) of the HiPER Project. These experiments concern the study of the physics connected to "Advanced Ignition Schemes", i.e. the Fast Ignition and the Shock Ignition Approaches to Inertial Fusion. Such schemes are aimed at achieving a higher gain, as compared to the classical approach which is used in NIF, as required for future reactors, and making fusion possible with smaller facilities. In particular, a series of experiments related to Fast Ignition were performed at the RAL (UK) and LULI, France) Laboratories and were addressed to study the propagation of fast electrons (created by a short-pulse ultra-high-intensity beam) in compressed matter, created either by cylindrical implosions or by compression of planar targets by (planar) laser-driven shock waves. A more recent experiment was performed at PALS and investigated the laser-plasma coupling in the 1016 W/cm2 intensity regime of interest for Shock Ignition.
Popa, L M; Repanovici, R; Iliescu, R
A study was conducted on the protective effect of some ribosomal preparations, isolated from chorionic-allantoic membranes of chicken embryos, infected or not with parainfluenza (Sendai) or influenza (AoPR8) virus, in mice experimentally inoculated with influenza virus strain AoPR8 adapted to the mouse. Results showed that the tested preparation, containing ribosomes and polysomes isolated from chorio-allantoic membranes of Sendai virus inoculated chicken embryos, ensure the mice complete protection against AoPR8 virus, if administrated before the control infection.
A method has been devised for inhibiting the normal enteric flora, permitting long term asymptomatic enteric infections of mice and guinea pigs with streptomycin-resistant strains of Shigella flexneri or Vibrio cholerae. Introduction of a streptomycin-resistant strain of E. coli into the intestinal tract of experimental animals resulted in a rapid elimination of the enteric pathogens studied. No in vitro production of antibiotic substances by this coli strain could be demonstrated. Active and oral passive immunization did not noticeably influence the number of Shigella or Vibrio organisms recoverable from the feces of infected animals. PMID:13357693
Castro-Sesquen, Yagahira E.; Gilman, Robert H.; Yauri, Verónica; Angulo, Noelia; Verastegui, Manuela; Velásquez, Daniel E.; Sterling, Charles R.; Martin, Diana; Bern, Caryn
The guinea pig (Cavia porcellus) is a natural reservoir for Trypanosoma cruzi but has seldom been used as an experimental infection model. We developed a guinea pig infection model for acute and chronic Chagas disease. Seventy-two guinea pigs were inoculated intradermally with 104 trypomastigotes of T. cruzi strain Y (experimental group); 18 guinea pigs were used as control group. Eight animals from the experimental group and two from the control group were sacrificed 5, 15, 20, 25, 40, 55, 115, 165, and 365 days after inoculation. During the acute phase (15 to 55 days), we observed parasitemia (with a peak on day 20) and positive IgM and IgG Western blots with anti-shed acute-phase antigen bands. The cardiac tissue showed vasculitis, necrosis (on days 40 to 55), moderate to severe inflammation, and abundant amastigote nests. Smaller numbers of amastigote nests were also present in kidney, brain, and other organs. In the early chronic phase (115 to 165 days), parasitemia disappeared and anti–T. cruzi IgG antibodies were still detectable. In cardiac tissue, the number of amastigote nests and the grade of inflammation decreased. In the chronic phase (365 days), the cardiac tissue showed vasculitis and fibrosis; detectable parasite DNA was associated with higher grades of inflammation. The experimental T. cruzi infection model in guinea pigs shows kinetics and pathologic changes similar to those of the human disease. PMID:21703410
Nemeth, N.M.; Hahn, D.C.; Gould, D.H.; Bowen, R.A.
Eastern Screech Owls (EASOs) were experimentally infected with the pathogenic New York 1999 strain of West Nile virus (WNV) by subcutaneous injection or per os. Two of nine subcutaneously inoculated birds died or were euthanatized on 8 or 9 days postinfection (DPI) after <24 hr of lethargy and recumbency. All subcutaneously inoculated birds developed levels of viremia that are likely infectious to mosquitoes, with peak viremia levels ranging from 105.0 to 109.6 plaque-forming units/ml. Despite the viremia, the remaining seven birds did not display signs of illness. All birds alive beyond 5 DPI seroconverted, although the morbid birds demonstrated significantly lower antibody titers than the clinically normal birds. Cagemates of infected birds did not become infected. One of five orally exposed EASOs became viremic and seroconverted, whereas WNV infection in the remaining four birds was not evident. All infected birds shed virus via the oral and cloacal route. Early during infection, WNV targeted skin, spleen, esophagus, and skeletal muscle. The two morbid owls had myocardial and skeletal muscle necrosis and mild encephalitis and nephritis, whereas some of the clinically healthy birds that were sacrificed on 14 DPI had myocardial arteritis and renal phlebitis. WNV is a significant pathogen of EASOs, causing pathologic lesions with varying clinical outcomes.
Novacco, Marilisa; Wolf-Jäckel, Godelind; Riond, Barbara; Hofmann-Lehmann, Regina
'Candidatus Mycoplasma turicensis' is a feline hemoplasma species that was isolated in a cat with hemolytic anemia. PCR has been widely used to investigate and diagnose 'Candidatus Mycoplasma turicensis' infection, but so far, little is known about the humoral immune response in infected cats. Recently, enzyme-linked immunosorbent assays (ELISA) were developed to monitor anti-feline hemoplasma antibodies. The aim of the present study was to investigate the humoral immune response in cats experimentally infected with 'Candidatus Mycoplasma turicensis' and to monitor the influence of the pre-administration of methylprednisolone and subsequent antibiotic treatment. Serum and plasma samples from 15 specified pathogen-free cats infected with 'Candidatus Mycoplasma turicensis' were analyzed by ELISA. Seroconversion was demonstrated in all cats, and the antibodies remained detectable until the end of the study (up to 100 weeks post-exposure). In some cats, the ELISA seemed more sensitive and better able to demonstrate exposure to 'Candidatus Mycoplasma turicensis' than PCR. The peak antibody level occurred after the peak of the bacterial blood loads. The methylprednisolone administrations were associated with increased antibody levels, while antibiotic treatment, particularly with doxycycline, resulted in a decrease in antibody levels. Additionally, preliminary data indicated that three of four seropositive cats were protected from bacteremia after a subsequent challenge. In conclusion, the ELISA was found to be a useful tool to investigate the humoral immune response in hemoplasma-infected cats and a desirable addition to PCR to study the pathogenesis of hemoplasma infections.
Munhoz, Alexandre Dias; Mineo, Tiago Wilson Patriarca; Alessi, Antonio Carlos; Lopes, Carlos Wilson Gomes; Machado, Rosangela Zacarias
The aim of this study was to evaluate parasitism kinetics and tissue lesions in the first week of infection by Neospora caninum in dogs fed Gallus gallus chorioallantoic membranes (CMs) previously infected in ovo. Five two-month-old pups were used. Each dog was given five CMs that were previously infected with N. caninum via the oral route. Four animals were euthanized in the first week of infection. All four dogs had their stools examined one week prior to and up to the day they were euthanized. The stools of the uneuthanized dog were collected for 30 days. After euthanasia, organ sections were utilized for histopathology, immunohistochemistry, indirect immunofluorescent tissue reactions, PCR and real-time PCR to detect parasites. Necropsy revealed that the small and large intestines, spleen, and lungs were affected. No oocysts or N. caninum DNA were identified in the stool samples. Real-time PCR was the most sensitive technique used to detect the protozoa in tissues, which were identified in 41% of the analyzed samples. Our results indicate that an experimental model using previously infected CMs appears to be a useful model for the study of the host-parasite relationship during the infection's acute phase.
Elwakil, Hala S; Ali, Nehad M; Talaat, Roba M; Osman, Wesam M
Experimental duel infection with S. mansoni and E. granulosus was induced in mice to determine their effect on serum nitric oxide (NO) level and accordingly on the sequences of histopathological lesions affecting the liver. The results showed that serum NO level was significantly increased (p<0.05) in mice infected with both parasites (GI) in comparison to either S. mansoni (GIV) or E. granulosus (GV). The NO elevation on hepatic pathological lesions of both diseases showed a marked reduction of granuloma size with absence of concentric fibrosis in GI as early as 4 weeks of concomitant infection as compared to GIV. In spite of the significant increase of NO level when E. granulosus infection induced in late stages of schistosomisais (GsII & III), yet granuloma size was not suppressed. Also, there was absence or death of hydatid cyst in mice (GI) compared to E. granulosus (GV). So, the duel infection with the two parasites affected serum NO level and hepatic histopathology, by ameliorative or deteriorative effects, according to duration of infection with either.
Dalgaard, Tina S; Skovgaard, Kerstin; Norup, Liselotte R; Pleidrup, Janne; Permin, Anders; Schou, Torben W; Vadekær, Dorte F; Jungersen, Gregers; Juul-Madsen, Helle R
Ascaridia galli is a gastrointestinal nematode infecting chickens. Chickens kept in alternative rearing systems or at free-range experience increased risk for infection with resulting high prevalences. A. galli infection causes reduced weight gain, decreased egg production and in severe cases increased mortality. More importantly, the parasitised chickens are more susceptible to secondary infections and their ability to develop vaccine-induced protective immunity against other diseases may be compromised. Detailed information about the immune response to the natural infection may be exploited to enable future vaccine development. In the present study, expression of immune genes in the chicken spleen during an experimental infection with A. galli was investigated using the Fluidigm(®) BioMark™ microfluidic qPCR platform which combines automatic high-throughput with attractive low sample and reagent consumption. Spleenic transcription of immunological genes was compared between infected chickens and non-infected controls at week 2, 6, and 9 p.i. corresponding to different stages of parasite development/maturation. At week 2 p.i. increased expression of IL-13 was observed in infected chickens. Increased expression of MBL, CRP, IFN-α, IL-1β, IL-8, IL-12β and IL-18 followed at week 6 p.i. and at both week 6 and 9 p.i. expression of DEFβ1 was highly increased in infected chickens. In summary, apart from also earlier reported increased expression of the Th2 signature cytokine IL-13 we observed only few differentially expressed genes at week 2 p.i. which corresponds to the larvae histotrophic phase. In contrast, we observed increased expression of pro-inflammatory cytokines and acute phase proteins in infected chickens, by week 6 p.i. where the larvae re-enter the intestinal lumen. Increased expression of DEFβ1 was observed in infected chickens at week 6 p.i. but also at week 9 p.i. which corresponds to a matured stage where adult worms are present in the
Gasbarrini, G; Pretolani, S; Bonvicini, F; Gatto, M R; Tonelli, E; Mégraud, F; Mayo, K; Ghironzi, G; Giulianelli, G; Grassi, M
Helicobacter pylori is present worldwide but few large population studies exist on the epidemiology of the infection. A random cross sectional study was performed of H pylori infection in the adult population of San Marino, a European country with high gastric cancer rate, to assess its prevalence and to evaluate its relations with gastrointestinal disease. In 2237 subjects (77% of the initial sample) H pylori IgG antibodies were detected with enzyme linked immunosorbent assay (ELISA) and immunoblotting. A questionnaire including questions about occupation, place of birth, and smoking was given to all subjects. Dyspepsia, peptic ulcer, and gastric cancer in the subjects, relatives, and partners as well as use of drug, dental treatment/prostheses, and gastrointestinal endoscopies, were evaluated by multivariate analysis. H pylori prevalence was of 51%, increased with age from 23% (20-29 years) to 68% (> or = 70 years), and was higher among manual workers. H pylori was independently associated with ulcer (OR = 1.63, 95% confidence intervals (CI) = 1.16 to 2.27), H2 antagonists (OR = 1.94, 95% CI = 1.21 to 3.10), and benzodiazepines (OR = 1.57, 95% CI = 1.02 to 2.42), dental prostheses (OR = 1.25, 95% CI = 1.05 to 1.49), gastroscopy in the past five years (OR = 1.50, 95% CI = 1.05 to 2.14), peptic ulcer in siblings (OR = 1.52, 95% CI = 1.09 to 2.12), gastric cancer in father (OR = 1.61, 95% CI = 1.02 to 2.52). The association of seropositivity with history of ulcer, gastric cancer in family, gastroscopy, and H2 antagonists suggests that H pylori is an epidemiological key factor in the pathogenesis of gastroduodenal diseases in this area. PMID:7615270
Mendlovic, Fela; Cruz-Rivera, Mayra; Ávila, Guillermina; Vaughan, Gilberto; Flisser, Ana
Taenia solium causes two diseases in humans, cysticercosis and taeniosis. Tapeworm carriers are the main risk factor for neurocysticercosis. Limited information is available about the immune response elicited by the adult parasite, particularly the induction of Th2 responses, frequently associated to helminth infections. Calreticulin is a ubiquitous, multifunctional protein involved in cellular calcium homeostasis, which has been suggested to play a role in the regulation of immune responses. In this work, we assessed the effect of recombinant T. solium calreticulin (rTsCRT) on the cytokine, humoral and cellular responses upon experimental infection in Syrian Golden hamsters (Mesocricetus auratus). Animals were infected with T. solium cysticerci and euthanized at different times after infection. Specific serum antibodies, proliferative responses in mesenteric lymph nodes and spleen cells, as well as cytokines messenger RNA (mRNA) were analyzed. The results showed that one third of the infected animals elicited anti-rTsCRT IgG antibodies. Interestingly, mesenteric lymph node (MLN) cells from either infected or non-infected animals did not proliferate upon in vitro stimulation with rTsCRT. Additionally, stimulation with a tapeworm crude extract resulted in increased expression of IL-4 and IL-5 mRNA. Upon stimulation, rTsCRT increased the expression levels of IL-10 in spleen and MLN cells from uninfected and infected hamsters. The results showed that rTsCRT favors a Th2-biased immune response characterized by the induction of IL-10 in mucosal and systemic lymphoid organs. Here we provide the first data on the cytokine, antibody and cellular responses to rTsCRT upon in vitro stimulation during taeniasis.
Mendlovic, Fela; Cruz-Rivera, Mayra; Ávila, Guillermina; Vaughan, Gilberto; Flisser, Ana
Taenia solium causes two diseases in humans, cysticercosis and taeniosis. Tapeworm carriers are the main risk factor for neurocysticercosis. Limited information is available about the immune response elicited by the adult parasite, particularly the induction of Th2 responses, frequently associated to helminth infections. Calreticulin is a ubiquitous, multifunctional protein involved in cellular calcium homeostasis, which has been suggested to play a role in the regulation of immune responses. In this work, we assessed the effect of recombinant T. solium calreticulin (rTsCRT) on the cytokine, humoral and cellular responses upon experimental infection in Syrian Golden hamsters (Mesocricetus auratus). Animals were infected with T. solium cysticerci and euthanized at different times after infection. Specific serum antibodies, proliferative responses in mesenteric lymph nodes and spleen cells, as well as cytokines messenger RNA (mRNA) were analyzed. The results showed that one third of the infected animals elicited anti-rTsCRT IgG antibodies. Interestingly, mesenteric lymph node (MLN) cells from either infected or non-infected animals did not proliferate upon in vitro stimulation with rTsCRT. Additionally, stimulation with a tapeworm crude extract resulted in increased expression of IL-4 and IL-5 mRNA. Upon stimulation, rTsCRT increased the expression levels of IL-10 in spleen and MLN cells from uninfected and infected hamsters. The results showed that rTsCRT favors a Th2-biased immune response characterized by the induction of IL-10 in mucosal and systemic lymphoid organs. Here we provide the first data on the cytokine, antibody and cellular responses to rTsCRT upon in vitro stimulation during taeniasis. PMID:25811778
Lankton, Julia; Werner, Katrien; Falendysz, Elizabeth A.; McCurley, Kevin; Blehert, David S.
ABSTRACT Snake fungal disease (SFD) is an emerging skin infection of wild snakes in eastern North America. The fungus Ophidiomyces ophiodiicola is frequently associated with the skin lesions that are characteristic of SFD, but a causal relationship between the fungus and the disease has not been established. We experimentally infected captive-bred corn snakes (Pantherophis guttatus) in the laboratory with pure cultures of O. ophiodiicola. All snakes in the infected group (n = 8) developed gross and microscopic lesions identical to those observed in wild snakes with SFD; snakes in the control group (n = 7) did not develop skin infections. Furthermore, the same strain of O. ophiodiicola used to inoculate snakes was recovered from lesions of all animals in the infected group, but no fungi were isolated from individuals in the control group. Monitoring progression of lesions throughout the experiment captured a range of presentations of SFD that have been described in wild snakes. The host response to the infection included marked recruitment of granulocytes to sites of fungal invasion, increased frequency of molting, and abnormal behaviors, such as anorexia and resting in conspicuous areas of enclosures. While these responses may help snakes to fight infection, they could also impact host fitness and may contribute to mortality in wild snakes with chronic O. ophiodiicola infection. This work provides a basis for understanding the pathogenicity of O. ophiodiicola and the ecology of SFD by using a model system that incorporates a host species that is easy to procure and maintain in the laboratory. PMID:26578676
Cousins, Scott W; Espinosa-Heidmann, Diego G; Miller, Daniel M; Pereira-Simon, Simone; Hernandez, Eleut P; Chien, Hsin; Meier-Jewett, Courtney; Dix, Richard D
The neovascular (wet) form of age-related macular degeneration (AMD) leads to vision loss due to choroidal neovascularization (CNV). Since macrophages are important in CNV development, and cytomegalovirus (CMV)-specific IgG serum titers in patients with wet AMD are elevated, we hypothesized that chronic CMV infection contributes to wet AMD, possibly by pro-angiogenic macrophage activation. This hypothesis was tested using an established mouse model of experimental CNV. At 6 days, 6 weeks, or 12 weeks after infection with murine CMV (MCMV), laser-induced CNV was performed, and CNV severity was determined 4 weeks later by analysis of choroidal flatmounts. Although all MCMV-infected mice exhibited more severe CNV when compared with control mice, the most severe CNV developed in mice with chronic infection, a time when MCMV-specific gene sequences could not be detected within choroidal tissues. Splenic macrophages collected from mice with chronic MCMV infection, however, expressed significantly greater levels of TNF-α, COX-2, MMP-9, and, most significantly, VEGF transcripts by quantitative RT-PCR assay when compared to splenic macrophages from control mice. Direct MCMV infection of monolayers of IC-21 mouse macrophages confirmed significant stimulation of VEGF mRNA and VEGF protein as determined by quantitative RT-PCR assay, ELISA, and immunostaining. Stimulation of VEGF production in vivo and in vitro was sensitive to the antiviral ganciclovir. These studies suggest that chronic CMV infection may serve as a heretofore unrecognized risk factor in the pathogenesis of wet AMD. One mechanism by which chronic CMV infection might promote increased CNV severity is via stimulation of macrophages to make pro-angiogenic factors (VEGF), an outcome that requires active virus replication.
Scorza, Andrea V; Radecki, Steven V; Lappin, Michael R
This study evaluated the effect of two combination products containing febantel, pyrantel, and praziquantel (FPP) for the treatment of Giardia species in experimentally infected kittens. In experiment 1, five kittens were administered the United States (US) formulation of FPP at doses of 37.8 mg/kg, 7.56 mg/kg, and 7.56 mg/kg, respectively, PO, q24h, for 5 days and four kittens remained as controls. In experiment 2, five kittens were administered the European formulation of FPP at the doses of 12.5 mg/kg, 12 mg/kg, and 4.16 mg/kg, respectively, PO, q24h, for 5 days and four kittens remained as controls. In experiment 3, six kittens were administered the US formulation of FPP at 56.5 mg/kg, 11.3 mg/kg, 11.3 mg/kg, respectively, PO, q24h, for 5 days and five kittens remained as controls. Thirteen days after treatment, kittens testing negative for Giardia species cysts were administered 20 mg/kg methylprednisolone acetate, IM, weekly for a maximum of two injections. Feces were analyzed for Giardia species cysts using a direct immunofluorescence test. After experiment 3, four of the six treated kittens, but no control kittens, remained negative for Giardia species after the administration of methylprednisolone acetate.
Gabriël, S; Johansen, M V; Pozio, E; Smit, G S A; Devleesschauwer, B; Allepuz, A; Papadopoulos, E; van der Giessen, J; Dorny, P
Taenia solium taeniasis/cysticercosis is a neglected zoonotic disease complex occurring primarily in developing countries. Though claimed eradicated from the European Union (EU), an increasing number of human neurocysticercosis cases is being detected. Risk factors such as human migration and movement of pigs/pork, as well as the increasing trend in pig rearing with outside access are discussed in this review. The entry of a tapeworm carrier into the EU seems a lot more plausible than the import of infected pork. The establishment of local transmission in the EU is presently very unlikely. However, considering the potential changes in risk factors, such as the increasing trend in pig farming with outdoor access, the increasing human migration from endemic areas into the EU, this situation might change, warranting the establishment of an early warning system, which should include disease notification of taeniasis/cysticercosis both in human and animal hosts. As currently human-to-human transmission is the highest risk, prevention strategies should focus on the early detection and treatment of tapeworm carriers, and should be designed in a concerted way, across the EU and across the different sectors.
Crobach, M J T; Planche, T; Eckert, C; Barbut, F; Terveer, E M; Dekkers, O M; Wilcox, M H; Kuijper, E J
In 2009 the first European Society of Clinical Microbiology and Infectious Diseases (ESCMID) guideline for diagnosing Clostridium difficile infection (CDI) was launched. Since then newer tests for diagnosing CDI have become available, especially nucleic acid amplification tests. The main objectives of this update of the guidance document are to summarize the currently available evidence concerning laboratory diagnosis of CDI and to formulate and revise recommendations to optimize CDI testing. This update is essential to improve the diagnosis of CDI and to improve uniformity in CDI diagnosis for surveillance purposes among Europe. An electronic search for literature concerning the laboratory diagnosis of CDI was performed. Studies evaluating a commercial laboratory test compared to a reference test were also included in a meta-analysis. The commercial tests that were evaluated included enzyme immunoassays (EIAs) detecting glutamate dehydrogenase, EIAs detecting toxins A and B and nucleic acid amplification tests. Recommendations were formulated by an executive committee, and the strength of recommendations and quality of evidence were graded using the Grades of Recommendation Assessment, Development and Evaluation (GRADE) system. No single commercial test can be used as a stand-alone test for diagnosing CDI as a result of inadequate positive predictive values at low CDI prevalence. Therefore, the use of a two-step algorithm is recommended. Samples without free toxin detected by toxins A and B EIA but with positive glutamate dehydrogenase EIA, nucleic acid amplification test or toxigenic culture results need clinical evaluation to discern CDI from asymptomatic carriage.
Hughes, David J; Kipar, Anja; Leeming, Gail; Sample, Jeffery T; Stewart, James P
MuHV-4 is a natural pathogen of rodents of the genus Apodemus (e.g., wood mice, yellow-necked mice) and Myodes glareolus (bank voles). We report experimental MuHV-4 infection of bank voles in comparison with infection of A. sylvaticus (wood mice) and BALB/c mice. Like in wood mice, the level of productive replication in the lungs of bank voles was significantly lower than in BALB/c mice. In contrast to other hosts, however, the level of latent infection in the lung and spleen of bank voles was extremely low. These findings, together with those of previous studies, suggest that bank voles are an occasional and inefficient host for MuHV-4.
Malafaia, Guilherme; Talvani, André
This paper reviews the scientific knowledge about protein-energy and micronutrient malnutrition in the context of Chagas disease, especially in experimental models. The search of articles was conducted using the electronic databases of SciELO (Scientific Electronic Library Online), PubMed and MEDLINE published between 1960 and March 2010. It was possible to verify that nutritional deficiencies (protein-energy malnutrition and micronutrient malnutrition) exert a direct effect on the infection by T. cruzi. However, little is known about the immunological mechanisms involved in the relationship “nutritional deficiencies and infection by T. cruzi”. A hundred years after the discovery of Chagas disease many aspects of this illness still require clarification, including the effects of nutritional deficiencies on immune and pathological mechanisms of T. cruzi infection. PMID:21577255
Choi, Chang-Jin; Lee, Hye-Jung; Go, Jai-Hyang; Park, Yun-Kyu; Chai, Jong-Yil; Seo, Min
Reptiles were known to serve as paratenic hosts for Centrorhynchus (Acanthocephala: Centrorhynchidae) in Korea, but the infection course in experimental animals was not elucidated yet. In this study, the tiger keelback snakes (Rhabdophis tigrinus) were collected and digested with artificial pepsin solution, and the larvae of Centrorhynchus were recovered from them. Then, the collected larvae were orally infected to rats for developmental observations. In rats, all the larvae were observed outside the intestine on day 3 post-infection (PI), including the mesentery and abdominal muscles. As for the development in rats, the ovary of Centrorhynchus sp. was observed at day 15 PI, and the cement glands were 3 in number. Based on the morphological characteristics, including the arrangement of proboscis hooks, these larvae proved to be a species of Centrorhynchus, and more studies were needed for species identification.
Geleta, J.N.; Shimoda, W.; Mercer, H.D.
The excretion rate of (3H)prednisolone from clinically normal and experimentally infected udders of 10 lactating cows was studied. Each quarter of 6 cows was injected with a single dose of (3H)prednisolone mixed with non-radioactive prednisolone equivalent to 10 mg in 10 ml of peanut oil base. Each of the remaining 4 cows was given 40 mg of nonradioactive prednisolone and (3H)prednisolone in 60% ethanol IV. Control and postadministration samples of blood, milk, and urine were examined for radioactivity. The effects of (3H)prednisolone were evaluated in the same cows, first in clinically normal udders, then 2 weeks later in udders experimentally infected with Streptococcus agalactiae. Absorption and elimination of prednisolone were the same before and after induced infection. Within 3 hours after intramammary injection, 95% of the labeled prednisolone was absorbed systemically, less than 5% of this dose was recovered in milk, and 29% was excreted in urine. After IV injection of (3H)prednisolone, less than 0.2% of the total radioactivity was recovered in milk and less than 46% was excreted in urine. Clinical mastitis induced by S agalactiae was moderate. Circulating blood leukocytes and somatic cells in the milk of normal cows remained essentially unchanged. The leukocyte response to induced infection was rapid in blood and milk. Large numbers of leukocytes were noticed in the milk and a severe leukopenia occurred. Prednisolone treatment did not alter the number of somatic cells in milk or reduce the inflammatory response of experimentally infected cows.
The present study was aimed at investigating the effect of experimental infection by Trypanosoma vivax in different stages of pregnancy, determining the pathogenesis of reproductive failure, and confirming transplacental transmission. We used 12 pregnant ewes distributed into four experimental groups: G1, was formed by three ewes infected with T. vivax in the first third of pregnancy (30 days); G2 comprised three infected ewes in the final third of pregnancy (100 days); G3 and G4 were composed of three non-infected ewes with the same gestational period, respectively. Each ewe of G1 and G2 was inoculated with 1.25 × 105 tripomastigotes. Clinical examination, determination of parasitemia, serum biochemistry (albumin, total protein, glucose, cholesterol, and urea), packed cell volume (PCV), serum progesterone, and pathological examination were performed. Placenta, amniotic fluid, blood and tissues from the fetuses and stillbirths were submitted to PCR. Two ewes of G1 (Ewe 1 and 3) presented severe infection and died in the 34th and 35th days post-infection (dpi), respectively; but both fetuses were recovered during necropsy. In G2, Ewe 5 aborted two fetuses on the 130th day (30 dpi) of pregnancy; and Ewe 6 aborted one fetus in the 140th day (40 dpi) of gestation. Ewes 2 and 4 delivered two weak lambs that died five days after birth. Factors possibly involved with the reproductive failure included high parasitemia, fever, low PCV, body score, serum glucose, total protein, cholesterol, and progesterone. Hepatitis, pericarditis, and encephalitis were observed in the aborted fetuses. The presence of T. vivax DNA in the placenta, amniotic fluid, blood, and tissues from the fetuses confirms the transplacental transmission of the parasite. Histological lesion in the fetuses and placenta also suggest the involvement of the parasite in the etiopathogenesis of reproductive failure in ewes. PMID:23289625
Gaze, Soraya; McSorley, Henry J; Daveson, James; Jones, Di; Bethony, Jeffrey M; Oliveira, Luciana M; Speare, Richard; McCarthy, James S; Engwerda, Christian R; Croese, John; Loukas, Alex
The mucosal cytokine response of healthy humans to parasitic helminths has never been reported. We investigated the systemic and mucosal cytokine responses to hookworm infection in experimentally infected, previously hookworm naive individuals from non-endemic areas. We collected both peripheral blood and duodenal biopsies to assess the systemic immune response, as well as the response at the site of adult worm establishment. Our results show that experimental hookworm infection leads to a strong systemic and mucosal Th2 (IL-4, IL-5, IL-9 and IL-13) and regulatory (IL-10 and TGF-β) response, with some evidence of a Th1 (IFN-γ and IL-2) response. Despite upregulation after patency of both IL-15 and ALDH1A2, a known Th17-inducing combination in inflammatory diseases, we saw no evidence of a Th17 (IL-17) response. Moreover, we observed strong suppression of mucosal IL-23 and upregulation of IL-22 during established hookworm infection, suggesting a potential mechanism by which Th17 responses are suppressed, and highlighting the potential that hookworms and their secreted proteins offer as therapeutics for human inflammatory diseases.
Vieira, Monica L.; de Andrade, Sonia A.; Morais, Zenaide M.; Vasconcellos, Silvio A.; Dagli, Maria Lucia Z.; Nascimento, Ana Lucia T. O.
Leptospirosis is a worldwide zoonotic and neglected infectious disease of human and veterinary concern, caused by pathogenic Leptospira species. Although bleeding is a common symptom of severe leptospirosis, the cause of hemorrhage is not completely understood. In severe infections, modulation of hemostasis by pathogens is an important virulence mechanism, and hemostatic impairments such as coagulation/fibrinolysis dysfunction are frequently observed. Here, we analyze the coagulation status of experimentally infected hamsters in an attempt to determine coagulation interferences and the origin of leptospirosis hemorrhagic symptomatology. Hamsters were experimentally infected with L. interrogans. The lungs, kidneys, and livers were collected for culture, histopathology, and coagulation assays. L. interrogans infection disturbs normal coagulation in the organs of animals. Our results suggest the presence of a thrombin-like factor or FX activator, which is able to activate FII in the leptospirosis organ extracts. The activity of those factors is accelerated in the prothrombinase complex. Additionally, we show for the first time that live leptospires act as a surface for the prothrombinase complex assembly. Our results contribute to the understanding of leptospirosis pathophysiological mechanisms and may open new routes for the discovery of novel treatments in the severe manifestations of the disease.
Afonso, Sónia M S; Neves, Luis; Pondja, Alberto; Macuamule, Cristiano; Mukaratirwa, Samson; Arboix, Margarita; Cristòfol, Carles; Capece, Bettencourt P S
A controlled trial was conducted to evaluate the efficacy of three therapeutics regimes of albendazole (ABZ) against Taenia multiceps larvae in experimental infected goats. Forty-nine goats experimentally infected with 3000 T. multiceps eggs were selected and randomly divided into treatment or control groups. Treatment with 10mg/kg for 3 days for group 1 (G1), 10mg/kg for group 2 (G2) and 20mg/kg/day for group 3 (G3) was applied 2 months after infection; group 4 (G4) served as a control group. A treatment with doses of 10mg/kg/day for 3 days on group 5 (G5) and group 6 (G6) was used as control, 5 months after the infection. The efficacy of ABZ was assessed as percentage of non-viable cysts which were determined by morphologic characteristics, movement and methyl blue staining technique. The efficacy of ABZ against 2 months old cysts was significantly different from the control and were 90.3% (28/31), 72.7% (8/11) and 73.9% (14/19) for G1, G2 and G3, respectively. No differences were observed in cyst viability between treated and control groups for 5-month old cysts. The results in this study indicate that ABZ is effective in goats against 2-month-old cysts of T. multiceps larva located in tissues outside the brain.
Hafez, Hafez Mohamed; Hauck, Rüdiger
Histomoniasis (infectious enterohepatitis, blackhead) is caused by the protozoan parasite Histomonas meleagridis (H. meleagridis). After the ban of all prophylactic and therapeutic drugs in the European Union, histomoniasis is increasingly responsible for considerable economic problems to the poultry industry. The aim of this study was to investigate the effect of a herbal product with extracts from cinnamon, garlic, lemon, and rosemary on H. meleagridis in turkey poults in vivo. For this purpose, 60 two-week-old poults were divided into three groups. Group 1 received the herbal product in the feed six days before infection and in water three days before infection, then in feed and drinking water until the end of the experiment. Groups 2 and 3 were left untreated. At week 3 of age, Groups 1 and 2 were infected intracloacally with H. meleagridis. Three weeks after infection the surviving birds were euthanized and examined for pathological lesions. Mortality was 20% in Group 1 and 50% in Group 2. There were no deaths in Group 3. DNA of histomonads was detected in all examined caeca and livers of the dead birds, but was not detected in any examined organ of the surviving birds of all groups. There was no noticeable difference in the lesion scores of the dead birds between the groups. The surviving birds of all groups did not show lesions post mortem. Since all effective prophylactic and therapeutic drugs against histomoniasis were banned in the EU, under given conditions the investigated herbal product seems to be an effective alternative for the reduction of mortality in turkeys caused by histomoniasis.
Finol, Héctor J.; Roschman-González, Antonio
The ultrastructural study in different tissues of mice experimentally infected with isolates of Trypanosoma evansi, Trypanosoma cruzi, and Leishmania mexicana reveals changes in cardiac myocytes, skeletal muscle fibers, and hepatic, adrenal, kidney, and spleen cells. Some of these changes were cytoarchitectural and others consisted of necrosis. Alterations in the microvasculature were also found. The mononuclear cell infiltrate included neutrophils, eosinophils, and macrophages. This work shows that diverse mice tissues are important target for trypanosomatids. PMID:25072046
AD-AL15 528 VIR61NIA UNIV CHARLOTTESVILLE DEPT OF DERMATOLOGY F/G 6/5 INVESTIGATIONS OF CROSS IMMUNITY BETWEEN LEISHMANIA TROPICA (JE--ETC(U) SEP 79...Investigations of Cross Immunity Between First Annual -- Leishmania tropica (Jericho) and Leishmania Feburary 1979-September 1979 braziliensis in... Leishmania tropica (Jericho) and LeisLmania braziliensis panamensis in Experimentally Infected Mystromys albacaudatus" First Annual Report Bruce E
Lopes, Francisco Canindé; de Paiva, Kaliane Alessandra Rodrigues; Coelho, Wesley Adson Costa; Nunes, Francisco Vítor Aires; da Silva, Jardel Bezerra; de Gouveia Mendes da Escóssia Pinheiro, Carolina; de Macêdo Praça, Layanne; Silva, Jean Berg Alves; Alves Freitas, Carlos Iberê; Batista, Jael Soares
The present study aimed to evaluate the effects of Trypanosoma vivax infection on the shape of the lactation curve and the milk quality of dairy goats experimentally infected with T. vivax. In total, twenty Saanen goats, aged 26-30 months and the same number of calving (two calvings), were divided into two experimental groups: an infected group, consisting of ten goats intravenously infected with 0.5 ml of blood containing approximately 1.25 × 10(5) trypomastigotes of T. vivax and ten uninfected animals as the control group. Clinical tests and hematocrit, parasitemia, and serum biochemistry evaluations were performed on all of the goats. Milk production was measured daily for 152 days by hand milking the goats and weighing the milk. Every seven days, physiochemical analyses were performed to evaluate the milk. Wood's nonlinear model was used to analyze the lactation curve parameters. The infected goats had high levels of parasitemia and hyperthermia, significantly reduced hematocrit, serum total protein, albumin, and glucose levels and increased cholesterol and urea concentrations. Wood's model indicated that the milk production of goats in the infected group declined sharply over a short period of time and produced a flattened yield curve and significant difference (P < 0.05) in the rate of increase of peak milk production, rate of decrease of milk production after the peak, day of peak milk production, and maximum peak milk production compared with that of the control group. Trypanosomiasis also affected the persistency of lactation, which was significantly reduced in goats in the infected group. In addition, the physico-chemical properties of the milk, including the fat content, defatted dry extracts (DDE) and protein content, decreased significantly (P < 0.05) in the goats in the infected group compared with those in the control group. The T. vivax-infected goats showed reduction in milk production, persistence of lactation, and fat levels, the
Sarcocystis neurona infections in raccoons (Procyon lotor): evidence for natural infection with sarcocysts, transmission of infection to opossums (Didelphis virginiana), and experimental induction of neurologic disease in raccoons.
Dubey, J P; Saville, W J; Stanek, J F; Lindsay, D S; Rosenthal, B M; Oglesbee, M J; Rosypal, A C; Njoku, C J; Stich, R W; Kwok, O C; Shen, S K; Hamir, A N; Reed, S M
Equine protozoal myeloencephalitis (EPM) is a serious neurologic disease of horses in the Americas and Sarcocystis neurona is the most common etiologic agent. The distribution of S. neurona infections follows the geographical distributions of its definitive hosts, opossums (Didelphis virginiana, Didelphis albiventris). Recently, cats and skunks were reported as experimental and armadillos as natural intermediate hosts of S. neurona. In the present report, raccoons (Procyon lotor) were identified as a natural intermediate host of S. neurona. Two laboratory-raised opossums were found to shed S. neurona-like sporocysts after ingesting tongues of naturally-infected raccoons. Interferon-gamma gene knockout (KO) mice fed raccoon-opossum-derived sporocysts developed neurologic signs. S. neurona was identified immunohistochemically in tissues of KO mice fed sporocysts and the parasite was isolated in cell cultures inoculated with infected KO mouse tissues. The DNA obtained from the tongue of a naturally-infected raccoon, brains of KO mice that had neurological signs, and from the organisms recovered in cell cultures inoculated with brains of neurologic KO mice, corresponded to that of S. neurona. Two raccoons fed mature S. neurona sarcocysts did not shed sporocysts in their feces, indicating raccoons are not likely to be its definitive host. Two raccoons fed sporocysts from opossum feces developed clinical illness and S. neurona-associated encephalomyelitis was found in raccoons killed 14 and 22 days after feeding sporocysts; schizonts and merozoites were seen in encephalitic lesions.
Watkins, M. L.; JET Team
The effectiveness of the pumped divertor during the 1994/95 experimental campaign of the Joint European Torus (JET) [P.-H. Rebut, R. J. Bickerton, and B. E. Keen, Nucl. Fusion 25, 1011 (1985)] has allowed the pursuit of a broad-based research program that is highly relevant to the International Thermonuclear Experimental Reactor (ITER) [K. Tomabechi and the ITER Team, Nucl. Fusion 31, 1135 (1991)]. High-performance hot-ion discharges with high confinement (H-modes) free of edge localized modes (ELMs) have set a JET record neutron rate in deuterium, but are limited by various magnetohydrodynamic (MHD) phenomena to βN<1.8, where βN=β/(I/aB), β is the ratio of the plasma pressure to the toroidal field pressure, I is the plasma current, B is the toroidal field, and a is the horizontal minor radius of the plasma. Quasi-steady-state ELMy H-modes have also been studied at high power, high current, and high β. The underlying energy transport exhibits a gyro-Bohm dependence that is lost close to the H-mode threshold and at high β. ELMy H-modes with detached divertor plasmas and radiative power exhaust (the operating regime foreseen for ITER) reduce the power loading to the targets, but at the expense of main plasma confinement and purity. Beryllium has been compared with carbon fiber composite as a divertor target material and melting has been induced at ITER reference off-normal heat loads, but only a moderate degree of self-protection of the beryllium target was found.
Singh, T Shantikumar; Sugiyama, Hiromu; Devi, K Ranjana; Singh, L Deben; Binchai, Sutheewan; Rangsiruji, Achariya
This study was aimed to find out the host-parasite relationship between Paragonimus heterotremus isolated as metacercariae from mountain crabs, Indochinamon manipurensis, in Manipur, India and laboratory animals such as puppies, albino rats, Swiss mice, guinea pigs, and rabbits, as experimental animals. The animals were fed with the metacercariae. Infected animals were sacrificed 35 to 430 days after feeding to recover worms, which were used to determine the developmental stages. Adult worms (n = 14) were recovered from 3 puppies > or = 70 days after feeding and immature worms (n = 25) were recovered from 2 other puppies 35 or 43 days after infection. The infection rate in puppies was 100%. Juvenile worms were recovered from 3 of 13 rats: 1 of 11 rats whose viscera and cavities were examined and both of two rats whose muscles were examined. Rats were not a suitable animal model for pulmonary infection with P. heterotremus. Mice, guinea pigs, and rabbits were also found to be insusceptible to pulmonary infection with P. heterotremus.
Da Silva, Aleksandro S.; Paim, Carlos Breno V.; França, Raqueli T.; Costa, Márcio M.; Duarte, Marta M. M. F.; Sangoi, Manuela B.; Moresco, Rafael N.; Monteiro, Silvia G.; Lopes, Sonia Terezinha A.
This study aimed to measure the levels of interferon-gamma (IFN-γ), tumor necrosis factor-alpha (TNF-α), interleukin 1 (IL-1), interleukin 6 (IL-6), and nitrite/nitrate (NOx) in serum of dogs experimentally infected with Rangelia vitalii. Twelve female mongrel dogs were divided into 2 groups; group A (uninfected controls) composed by healthy dogs (n=5) and group B consisting of dogs inoculated with R. vitalii (n=7). Animals were monitored by blood smear examinations, which showed intraerythrocytic forms of the parasite on day 5 post-infection (PI). Blood samples were collected through the jugular vein on days 0, 10, and 20 PI to determine the serum levels of IFN-γ, TNF-α, IL-1, IL-6, and NOx. Cytokines were assessed by ELISA quantitative sandwich technique, and NOx was measured by the modified Griess method. Cytokine levels (IFN-γ, TNF-α, IL-1, and IL-6) were increased (P<0.01) in serum of infected animals. Serum levels of NOx were also increased on days 10 PI (P<0.01) and 20 PI (P<0.05) in infected animals. Therefore, the infection with R. vitalii causes an increase in proinflammatory cytokines and nitric oxide content. These alterations may be associated with host immune protection against the parasite. PMID:23467990
de Azevedo, Maria Isabel; Ferreiro, Laerte; Da Silva, Aleksandro S; Tonin, Alexandre A; Monteiro, Danieli Urach; Casali, Emerson A; Moritz, Cesar E J; Schirmbeck, Gabriel H; Cardoso, Valesca V; Flores, Mariana M; Fighera, Rafael; Stefani, Lenita M; Santurio, Janio M
The present study was carried out to assess the participation of purines in the activation and modulation of inflammatory response of rats experimentally infected by Cryptococcus neoformans. Twenty four Wistar rats were divided into two groups of 12 animals each: Group A - uninfected control group and Group B - infected by C. neoformans. Blood was collected 20 and 50 days post-infection (PI) from six animals of each group in order to verify purine levels (adenosine triphosphate (ATP), adenosine diphosphate (ADP), adenosine monophosphate (AMP), adenosine (ADO), inosine (INO), hypoxanthine (HYPO), xanthine (XAN) and uric acid (URIC)). ATP levels were significantly increased (P < 0.05) in serum from infected animals on days 20 and 50 PI, as well as adenosine levels after 20 days PI on rats. On day 50 PI, levels of adenosine and uric acid were also reduced, but the levels of inosine and xanthine increased in animals infected by the fungus (P < 0.05). Therefore, it was possible to conclude that the purine levels in serum were altered and that these changes may be able to influence the pathogenesis of the disease caused by C. neoformans due the participation of purines (ATP and adenosine main) in the activation and modulation of inflammatory response.
Valiente Moro, C; Chauve, C; Zenner, L
Dermanyssus gallinae is an important ectoparasite of laying hens in Europe and it is suspected of being a vector of pathogens. We carried out an in vitro study to evaluate the role of D. gallinae as a vector of Salmonella enterica subsp. enterica serotype Enteritidis. Two means of infecting the mite were tested: through the blood meal and after cuticular contact. Mites became carriers of Salmonella immediately after the infection with 29% and 53%, respectively, for oral route and cuticular contact. This percentage increased over time until it reached 95% (D7) and 80% (D14). The numerical identification of bacteria on the selective medium SM ID demonstrated the multiplication of Salmonella inside previously infected mites. In addition, transovarial passage as well as transstadial passage (from N1 to N2 stages) were demonstrated. Moreover, the observation of a negative effect of Salmonella on Dermanyssus oviposition was also observed. Finally, previously infected mites were able to contaminate the blood during the blood meal. Therefore, it appears that D. gallinae may act as a biological vector of S. Enteritidis under experimental conditions. It may represent a suitable environment for the development of Salmonella and could be an additional factor for the persistence of salmonellosis infection between successive flocks.
Troke, P F; Andrews, R J; Brammer, K W; Marriott, M S; Richardson, K
UK-49,858 (fluconazole), a new, orally absorbed bis-triazole derivative, has been evaluated against systemic infections with Candida albicans in normal and immunosuppressed mice and against an intestinal infection with C. albicans in immunosuppressed mice. Orally administered ketoconazole was used as a comparison agent throughout, and orally administered amphotericin B was included for comparative in the experimental intestinal infection. In a 10-day dosage regimen, UK-49,858 was far more active than ketoconazole against systemic infections with C. albicans in normal and immunosuppressed mice. In normal mice, extension of UK-49,858 dosing to 30 days resulted in prolongation of survival to over 90 days, and up to 60% of treated animals had no detectable C. albicans in their kidneys. In addition, over 90% of mice with intestinal candidiasis had culture-negative feces after a 3-day treatment with UK-49,858, but only 62 and 23% of mice gave this response after amphotericin B and ketoconazole therapy, respectively. These data suggest that UK-49,858 may be of value in the treatment of systemic and gastrointestinal infections due to C. albicans in humans. PMID:3002246
Drummond, J.G.; Curtis, S.E.; Simon, J.; Norton, H.W.
Effects of atmospheric ammonia at 69.4 mg/m3 (100 ppm) on productive performance and respiratory tract health of young pigs (starting body weight averaged 7.5 kg) experimentally infected with Ascaris suum (50,000 embryonated ova administered by gavage when pigs were 5 weeks of age) were studied in 5 trials of 4 weeks each (when pigs were 5 to 9 weeks of age). Effects of atmospheric-ammonia exposure and ascarid infection on growth were additive. Compared with controls, percentage reductions in average daily gain were 32%, 28%, and 61% for ammonia-exposed, ascarid-infected, and combined ammonia plus ascarid groups, respectively. Ammonia exposure or ascarid infection alone depressed feed disappearance by 18%. Effects of the 2 factors were additive, resulting in a 35% reduction in feed disappearance. Pigs exposed to the combined factors had an average gain/feed ratio of 0.518, which was less than that of control pigs (0.546), but was greater than that of pigs exposed to atmospheric ammonia (0.489) or pigs infected with ascarids (0.501) alone. Liver scarring, due to larval migration, was not affected by ammonia exposure. Larval migration through the respiratory tract was not confirmed histopathologically in pigs killed 4 weeks after inoculation. A supplementary experiment was conducted which demonstrated that residual evidence of previous pulmonary larval migration was present 2 weeks after inoculation.
Angela Montesano, M.; Colley, Daniel G.; Eloi-Santos, Silvana; Freeman, George L.; Secor, W. Evan
Exposure to maternal idiotypes (Ids) or antigens might predispose a child to develop an immunoregulated, asymptomatic clinical presentation of schistosomiasis. We have used an experimental murine system to address the role of Ids in this immunoregulation. Sera from mice with 8-wk Schistosoma mansoni infection, chronic (20-wk infection) moderate splenomegaly syndrome (MSS), or chronic hypersplenomegaly syndrome (HSS) were passed over an S. mansoni soluble egg antigen (SEA) immunoaffinity column to prepare Ids (8WkId, MSS Id, HSS Id). Newborn mice were injected with 8WkId, MSS Id, HSS Id, or normal mouse immunoglobulin (NoMoIgG) and infected with S. mansoni 8 wk later. Mice exposed to 8WkId or MSS Id as newborns had prolonged survival and decreased morbidity compared with mice that received HSS Id or NoMoIgG. When stimulated with SEA, 8WkId, or MSS Id, spleen cells from mice neonatally injected with 8WkId or MSS Id produced more interferon γ than spleen cells from mice neonatally injected with HSS Id or NoMoIgG. Furthermore, neonatal exposure to 8WkId or MSS Id, but not NoMoIgG or HSS Id, led to significantly smaller granuloma size and lower hepatic fibrosis levels in infected mice. Together, these results indicate that perinatal exposure to appropriate anti-SEA Ids induces long-term effects on survival, pathology, and immune response patterns in mice subsequently infected with S. mansoni. PMID:9989978
Malone, Alyssa N.; Fletcher, Darcy M.; Vogt, Megan B.; Meyer, Stephen K.; Hess, Ann M.; Eckstein, Torsten M.
Johne’s disease is an infectious chronic inflammatory bowel disease in ruminants. The key factor for the management of this disease is an early positive diagnosis. Unfortunately, most diagnostics detect animals with Johne’s disease in the clinical stage with positive serology and/or positive fecal cultures. However, for effective management of the disease within herds, it is important to detect infected animals as early as possible. This might only be possible with the help of parameters not specific for Johne’s disease but that give an early indication for chronic infections such as weight development. Here we report our findings on the development of total body weight and weight gain during the first six months of goats experimentally infected to induce Johne’s disease. Twenty dairy goat kids age 2 to 5 days were included in this study. Goats were divided into two groups: a negative control group and a positive infected group. The weight was obtained weekly throughout the study. Goats of the positive group were infected at the age of seven weeks. We detected significant changes in weight gain and total body weight as early as one week after infection. Differences are significant throughout the six month time period. Weight as a non-specific parameter should be used to monitor infection especially in studies on Johne’s disease using the goat model. Our study suggests that goats with Johne’s disease have a reduced weight gain and reduced weight when compared with healthy goats of the same age. PMID:24349564
Oryan, A; Sadjjani, S M; Azizi, S
Toxocara cati (T. cati) and Toxocara canis (T. canis), roundworms of cats and dogs, are zoonotic parasites that cause visceral and ocular larval migrans in human beings. Humans and other paratenic hosts are infected by ingesting the infective Toxocara eggs from contaminated soil, unwashed hands, contaminated raw vegetables or ingestion of under-cooked organs and muscle tissues of infected paratenic hosts such as chickens, cattle and sheep. It has been shown that the seroprevalence of toxocariasis in the rural and urban children of southern Iran is high and more than 50% of cats of this area are also infected with T. cati. It is stated that consumption of raw chicken meat resulted in visceral toxocariasis. It is possible that poultry reared outdoors and feeding in open range system, gain Toxocara eggs from soil and or by eating infected earthworms as paratenic host. The aim of the present study was to investigate the effects of albendazole and febendazole in experimentally infected chickens with eggs of T. cati by histopathological and digestive methods. Pathologic lesions were observed only in the untreated group and larvae were detected in brain of 3 chickens of this group by squash method. No larva was observed at histopathological level in liver, lungs, brain, cardiac and skeletal muscles and other examined organs of either treated or untreated animals. No lesion was seen in other tissues of the infected untreated chickens. Treatment resulted in disappearance of the larvae and disappearance of the gross and histopathologic abnormalities from their organs. No detectable difference was observed in chemosusceptibility of the two drugs.
Schabuss, M; Gemeiner, M; Gleiss, A; Lewis, J W; Miller, I; Möstl, E; Schober, U; Tschulenk, W; Walter, I; Grillitsch, B
European chub Leuciscus cephalus collected from five localities in the lowland and subalpine regions of Austria were analysed for oestrogenic effects of endocrine-disrupting chemicals and the presence of the plerocercoid of the tapeworm Ligula intestinalis. Of 1494 chub analysed, only seven (six males, one female) were found to be infected with single, but large plerocercoids up to 15 cm in length. Ligula-infected fish showed comparatively immature gonads, as demonstrated by the gonadosomatic index and gamete developmental stages. Plasma levels of the egg precursor protein vitellogenin also showed concentrations ranging below the detection limit. The present results indicate that chub infected with L. intestinalis and exposed to exogenous oestrogenic compounds can result in reduced gonadal maturation and produce false oestrogen-positive diagnoses in male fish. For plasma vitellogenin levels, L. intestinalis infections can result in false oestrogen-negative diagnoses in male and female fish.
Brito, Luciana da S.; Pereira, Elder N.; da Silva, Augusta A.; Bentivóglio Costa Silva, Vinícius; Freitas, Fagner L. da C.
Through this study we assessed the metabolic and pathological changes in broilers experimentally infected with oocysts of Eimeria maxima. To perform the experiment, we used 150 broiler strain cooB males, with ten days of age, were randomized according to weight and randomly assigned to two experimental groups: the control group was inoculated with 0.5 mL of distilled water; the infected group inoculated with 0.5 mL of solution containing 5 × 104 sporulated oocysts of Eimeria maxima. The live performance was evaluated on day 0 (day of inoculation), 5°, 10°, 15°, 25°, and 35° dpi, being slaughtered by cervical dislocation, fifteen birds/group. Although the sum in meat production was higher in the control group, the weight of the heart and gizzard of the experimental animals showed no significant difference, while the liver had difference on day 5°, 15°, and 35° dpi. The pathologic evaluation showed congested mucosa and presence of large amounts of mucus at 6 dpi. Therefore, it is concluded that the dose of 5 × 104 E. maxima inoculated in the experimental group was enough to cause harm to the animal organism. PMID:26464925
Chen, Eunice C.; Liu, Maria; Brasky, Kathleen M.; Lanford, Robert E.; Kelly, Kristi R.; Bales, Karen L.; Schnurr, David P.; Canfield, Don R.; Patterson, Jean L.; Chiu, Charles Y.
Adenoviruses are DNA viruses that infect a number of vertebrate hosts and are associated with both sporadic and epidemic disease in humans. We previously identified a novel adenovirus, titi monkey adenovirus (TMAdV), as the cause of a fulminant pneumonia outbreak in a colony of titi monkeys (Callicebus cupreus) at a national primate center in 2009. Serological evidence of infection by TMAdV was also found in a human researcher at the facility and household family member, raising concerns for potential cross-species transmission of the virus. Here we present experimental evidence of cross-species TMAdV infection in common marmosets (Callithrix jacchus). Nasal inoculation of a cell cultured-adapted TMAdV strain into three marmosets produced an acute, mild respiratory illness characterized by low-grade fever, reduced activity, anorexia, and sneezing. An increase in virus-specific neutralization antibody titers accompanied the development of clinical signs. Although serially collected nasal swabs were positive for TMAdV for at least 8 days, all 3 infected marmosets spontaneously recovered by day 12 post-inoculation, and persistence of the virus in tissues could not be established. Thus, the pathogenesis of experimental inoculation of TMAdV in common marmosets resembled the mild, self-limiting respiratory infection typically seen in immunocompetent human hosts rather than the rapidly progressive, fatal pneumonia observed in 19 of 23 titi monkeys during the prior 2009 outbreak. These findings further establish the potential for adenovirus cross-species transmission and provide the basis for development of a monkey model useful for assessing the zoonotic potential of adenoviruses. PMID:23894316
Schnyder, Manuela; Fahrion, Anna; Riond, Barbara; Ossent, Pete; Webster, Pia; Kranjc, Asja; Glaus, Tony; Deplazes, Peter
The aim of this comparative study was to investigate the development of clinical signs and accompanying haematological, coproscopic and pathological findings as a basis for the monitoring of health condition of Angiostrongylus vasorum infected dogs. Six beagles were orally inoculated with 50 (n=3) or 500 (n=3) A. vasorum third stage larvae (L3) obtained from experimentally infected Biomphalaria glabrata snails. Two dogs were treated with moxidectin/imidacloprid spot-on solution and two further dogs with an oral experimental compound 92 days post infection (dpi), and were necropsied 166 dpi. Two untreated control dogs were necropsied 97 dpi. Prepatency was 47-49 days. Dogs inoculated with 500 L3 exhibited earlier (from 42 dpi) and more severe respiratory signs. Clinical signs resolved 12 days after treatment and larval excretion stopped within 20 days in all four treated dogs. Upon necropsy, 10 and 170 adult worms were recovered from the untreated dogs inoculated with 50 and 500 L3, respectively. Adult worms were also found in two treated dogs, in the absence of L1 or eggs. Despite heavy A. vasorum infection load and severe pulmonary changes including vascular thrombosis, only mild haematological changes were observed. Eosinophilia was absent but the presence of plasma cells was observed. Neutrophilic leucocytes showed a transient increase but only after treatment. Signs for coagulopathies were slight; nevertheless coagulation parameters were inoculation dose dependent. Ten weeks after treatment pulmonary fibrosis was still present. Infections starting from 50 L3 of A. vasorum had a massive impact on lung tissues and therefore on the health of affected dogs, particularly after prepatency, although only mild haematological abnormalities were evident.
Oliveira, A. V.; Roque-Barreira, M. C.; Sartori, A.; Campos-Neto, A.; Rossi, M. A.
likely that they are implicated in the pathogenesis of the mesangial proliferative glomerulonephritis in hamsters experimentally infected with L donovani. The glomerular changes may also explain the loss of immunoglobulins in the urine and the consequent lowering of serum immunoglobulin levels. Images Figure 2 Figure 3 Figure 4 PMID:4025511
Yaghfoori, Saeed; Razmi, Gholam Reza; Mohri, Mehrdad; Razavizadeh, Ali Reza Taghavi; Movassaghi, Ahmad Reza
The malignant ovine theileriosis is caused by Theileria lestoquardi, which is highly pathogenic in sheep. Theileriosis involves different organs in ruminants, but the effect of the disease on the cardiovascular system is unclear. To understand the pathogenesis of T. lestoquardi on the cardiovascular system, Baluchi breed sheep were infected with the mentioned parasite by releasing unfed adults of Hyalomma anatolicum anatolicum, which were infected with T. lestoquardi. The infected sheep were clinically examined on days 0, 2, 5, 7, 10, 12, 14, 17, and 21, and the blood samples were collected for biochemical parameters measurement. At termination of the experiment, the infected sheep were euthanized and pathological examinations of heart tissue were conducted. During experimental infection of sheep with T. lestoquardi, activities of cardiac troponin I (cTnI), lactate dehydrogenase, and aspartate aminotransferase, were significantly increased (P˂0.05), while a conspicuous decrease (P˂0.05) was observed in creatine phosphokinase activities. Alterations made in biochemical factors almost coincided with the presence of piroplasm in the blood and schizont in lymph nodes. Maximum and minimum of parasitemia in the sheep stood between 3.3% and 0.28%, respectively. In addition, electrocardiography revealed sinus tachycardia, sinus arrhythmia, sino-atrial block and ST-elevation, atrial premature beat, and alteration in QRS and in T waves' amplitude. Heart histopathological examination showed hyperemia, infiltration of mononuclear inflammatory cells into interstitial tissue, endocarditis, and focal necrosis of cardiac muscle cells. In addition, in one of the sheep, definite occurrence of infarction was observed. The results indicate that T. lestoquardi infection has devastating pathological impacts on the cardiovascular system of sheep. Furthermore, measurement of the cTnI amount is a useful biochemical factor for diagnosis and for better understanding of the severity and
Salinas-Carmona, Mario C.; Pérez-Rivera, Isabel
An experimental model of infection with Nocardia brasiliensis, used as an example of a facultative intracellular pathogen, was tested. N. brasiliensis was injected into the rear foot pads of BALB/c mice to establish an infection. Within 30 days, infected animals developed a chronic actinomycetoma infection. Batch cultures of N. brasiliensis were used to purify P61, P38, and P24 antigens; P61 is a catalase, and P38 is a protease with strong caseinolytic activity. Active and passive immunizations of BALB/c mice with these three purified soluble antigens were studied. Protection was demonstrated for actively immunized mice. However, immunity lasted only 30 days. Other groups of immunized mice were bled at different times, and their sera were passively transferred to naive recipients that were then infected with N. brasiliensis. Sera collected 5, 6, and 7 days after donor immunization conferred complete, long-lasting protection. The protective effect of passive immunity decreased when sera were collected 2 weeks after donor immunization. However, neither the early sera (1-, 2-, and 3-day sera) nor the later sera (30- or 45-day sera) prevented the infection. Hyperimmune sera with the highest levels of immunoglobulin G (IgG) to N. brasiliensis antigens did not protect at all. The antigens tested induced two IgM peaks. The first peak was present 3 days after immunization but was not antigen specific and did not transfer protection. The second peak was evident 7 days after immunization, was an IgM response, was antigen specific, and conferred protection. This results clearly demonstrate that IgM antibodies protect the host against a facultative intracellular bacterium. PMID:15385456
Müller, Joachim; Aguado-Martinez, Adriana; Manser, Vera; Balmer, Vreni; Winzer, Pablo; Ritler, Dominic; Hostettler, Isabel; Arranz-Solís, David; Ortega-Mora, Luis; Hemphill, Andrew
The naphthoquinone buparvaquone is currently the only drug used against theileriosis. Here, the effects of buparvaquone were investigated in vitro and in an experimental mouse model for Neospora caninum infection. In 4-day proliferation assays, buparvaquone efficiently inhibited N. caninum tachyzoite replication (IC50 = 4.9 nM; IC100 = 100 nM). However, in the long term tachyzoites adapted and resumed proliferation in the presence of 100 nM buparvaquone after 20 days of cultivation. Parasiticidal activity was noted after 9 days of culture in 0.5 µM or 6 days in 1 µM buparvaquone. TEM of N. caninum infected fibroblasts treated with 1 µM buparvaquone showed that the drug acted rather slowly, and ultrastructural changes were evident only after 3–5 days of treatment, including severe alterations in the parasite cytoplasm, changes in the composition of the parasitophorous vacuole matrix and a diminished integrity of the vacuole membrane. Treatment of N. caninum infected mice with buparvaquone (100 mg/kg) either by intraperitoneal injection or gavage prevented neosporosis symptoms in 4 out of 6 mice in the intraperitoneally treated group, and in 6 out of 7 mice in the group receiving oral treatment. In the corresponding controls, all 6 mice injected intraperitoneally with corn oil alone died of acute neosporosis, and 4 out of 6 mice died in the orally treated control group. Assessment of infection intensities in the treatment groups showed that, compared to the drug treated groups, the controls showed a significantly higher parasite load in the lungs while cerebral parasite load was higher in the buparvaquone-treated groups. Thus, although buparvaquone did not eliminate the parasites infecting the CNS, the drug represents an interesting lead with the potential to eliminate, or at least diminish, fetal infection during pregnancy. PMID:25941626
Müller, Joachim; Aguado-Martinez, Adriana; Manser, Vera; Balmer, Vreni; Winzer, Pablo; Ritler, Dominic; Hostettler, Isabel; Arranz-Solís, David; Ortega-Mora, Luis; Hemphill, Andrew
The naphthoquinone buparvaquone is currently the only drug used against theileriosis. Here, the effects of buparvaquone were investigated in vitro and in an experimental mouse model for Neospora caninum infection. In 4-day proliferation assays, buparvaquone efficiently inhibited N. caninum tachyzoite replication (IC50 = 4.9 nM; IC100 = 100 nM). However, in the long term tachyzoites adapted and resumed proliferation in the presence of 100 nM buparvaquone after 20 days of cultivation. Parasiticidal activity was noted after 9 days of culture in 0.5 µM or 6 days in 1 µM buparvaquone. TEM of N. caninum infected fibroblasts treated with 1 µM buparvaquone showed that the drug acted rather slowly, and ultrastructural changes were evident only after 3-5 days of treatment, including severe alterations in the parasite cytoplasm, changes in the composition of the parasitophorous vacuole matrix and a diminished integrity of the vacuole membrane. Treatment of N. caninum infected mice with buparvaquone (100 mg/kg) either by intraperitoneal injection or gavage prevented neosporosis symptoms in 4 out of 6 mice in the intraperitoneally treated group, and in 6 out of 7 mice in the group receiving oral treatment. In the corresponding controls, all 6 mice injected intraperitoneally with corn oil alone died of acute neosporosis, and 4 out of 6 mice died in the orally treated control group. Assessment of infection intensities in the treatment groups showed that, compared to the drug treated groups, the controls showed a significantly higher parasite load in the lungs while cerebral parasite load was higher in the buparvaquone-treated groups. Thus, although buparvaquone did not eliminate the parasites infecting the CNS, the drug represents an interesting lead with the potential to eliminate, or at least diminish, fetal infection during pregnancy.
Del Cacho, Emilio; Gallego, Margarita; Lillehoj, Hyun Soon; Quilez, Joaquin; Lillehoj, Erik P; Sánchez-Acedo, Caridad
Avian coccidiosis is caused by Eimeria, a unicellular, apicomplexan protist which primarily infects intestinal epithelia resulting in nutrient malabsorption and reduced growth of commercial poultry. Vaccination of chickens with exosomes isolated from antigen presenting cells containing parasite antigens (Ags) represents a promising alternative strategy to control avian coccidiosis, but is restricted in its commercial application due to limitations on production scale-up for mass immunization programs. Here, we report the biochemical and physiologic characteristics of exosomes derived from serum of Eimeria tenella-infected chickens and their feasibility for inducing protective immunity to experimental coccidiosis. Exosomes isolated from the serum of E. tenella-infected chickens contained a subset of protein Ags found in the intact parasite. Serum-derived exosomes containing these E. tenella Ags localized to the intestine and spleen following intramuscular injection into naïve chickens. In vitro ELISPOT assays revealed increased numbers of IL-2-, IL-4-, IL-6-, and IFN-γ-secreting cells in the intestine and spleen of exosome-administered chickens, compared with vehicle controls. Pre-immunization of chickens with serum exosomes from E. tenella-infected chickens increased both body weight gain and feed conversion efficiency, and reduced both fecal parasite shedding and gut lesion scores following parasite infection, compared with vehicle controls. Finally, immunization with CD80(+) serum exosomes stimulated greater numbers of cytokine-producing cells, and higher levels of protective immunity to E. tenella infection, compared with CD80(-) exosomes. These results suggest the possibility of producing an effective, parasite-free vaccine against avian coccidiosis under field conditions using serum-derived CD80(+) exosomes containing parasite Ags.
Christie, Rachel D; Marcus, Emily C; Wagner, April M; Besselsen, David G
Hamster parvovirus (HaPV) was isolated 2 decades ago from hamsters with clinical signs similar to those induced in hamsters experimentally infected with other rodent parvoviruses. Genetically, HaPV is most closely related to mouse parvovirus (MPV), which induces subclinical infection in mice. A novel MPV strain, MPV3, was detected recently in naturally infected mice, and genomic sequence analysis indicates that MPV3 is almost identical to HaPV. The goal of the present studies was to examine the infectivity of HaPV in mice. Neonatal and weanling mice of several mouse strains were inoculated with HaPV. Tissues, excretions, and sera were harvested at 1, 2, 4, and 8 wk after inoculation and evaluated by quantitative PCR and serologic assays specific for HaPV. Quantitative PCR detected viral DNA quantities that greatly exceeded the quantity of virus in inocula in multiple tissues of infected mice. Seroconversion to both nonstructural and structural viral proteins was detected in most immunocompetent mice 2 or more weeks after inoculation with HaPV. In neonatal SCID mice, viral transcripts were detected in lymphoid tissues by RT-PCR and viral DNA was detected in feces by quantitative PCR at 8 wk after inoculation. No clinical signs, gross, or histologic lesions were observed. These findings are similar to those observed in mice infected with MPV. These data support the hypothesis that HaPV and MPV3 are likely variants of the same viral species, for which the mouse is the natural rodent host with rare interspecies transmission to the hamster. PMID:20412687
Moro, Claire Valiente; Fravalo, Philippe; Amelot, Michel; Chauve, Claude; Zenner, Lionel; Salvat, Gilles
The poultry red mite (Dermanyssus gallinae) is the most important and common ectoparasite of laying hens in Europe. This haematophagous mite has been experimentally demonstrated to be a vector of Salmonella Enteritidis by acquiring bacteria through the blood meal or cuticular contact. We have evaluated another route of infection by orally inoculating chicks with mites previously infected by S. Enteritidis. Two methods of infecting the mites were tested: mites contaminated by cuticular contact or during the blood meal. After the washing of mites with paraformaldehyde, groups of 10 Salmonella-contaminated mites were inoculated individually into 1-day-old chicks. The titre of the inoculum suspension was evaluated by crushing mites and followed by bacteriological counting. It was 3x10(4) colony-forming units/chick and 2.7x10(6) colony-forming units/chick, respectively, for cuticular contact and orally mediated contamination of mites. Each bird was found to be positive 12 days post-inoculation. Salmonella colonized the intestinal tracts and invaded the livers and spleens. The caecal content concentration reached a mean level of S. Enteritidis of 8.5x10(4) most probable number (MPN) Salmonella/g. This experiment demonstrated the ability of mites to orally infect 1-day-old chicks with subsequent colonization and multiplication of Salmonella. Consequently, mites infected by S. Enteritidis constitute potential reservoir hosts of this bacterium, allowing it to persist in the poultry house as a source of infection for newly introduced animals. If contaminated mites are found in poultry facilities, effective red mite control should be performed before new batches are introduced into the facility.
Christie, Rachel D; Marcus, Emily C; Wagner, April M; Besselsen, David G
Hamster parvovirus (HaPV) was isolated 2 decades ago from hamsters with clinical signs similar to those induced in hamsters experimentally infected with other rodent parvoviruses. Genetically, HaPV is most closely related to mouse parvovirus (MPV), which induces subclinical infection in mice. A novel MPV strain, MPV3, was detected recently in naturally infected mice, and genomic sequence analysis indicates that MPV3 is almost identical to HaPV. The goal of the present studies was to examine the infectivity of HaPV in mice. Neonatal and weanling mice of several mouse strains were inoculated with HaPV. Tissues, excretions, and sera were harvested at 1, 2, 4, and 8 wk after inoculation and evaluated by quantitative PCR and serologic assays specific for HaPV. Quantitative PCR detected viral DNA quantities that greatly exceeded the quantity of virus in inocula in multiple tissues of infected mice. Seroconversion to both nonstructural and structural viral proteins was detected in most immunocompetent mice 2 or more weeks after inoculation with HaPV. In neonatal SCID mice, viral transcripts were detected in lymphoid tissues by RT-PCR and viral DNA was detected in feces by quantitative PCR at 8 wk after inoculation. No clinical signs, gross, or histologic lesions were observed. These findings are similar to those observed in mice infected with MPV. These data support the hypothesis that HaPV and MPV3 are likely variants of the same viral species, for which the mouse is the natural rodent host with rare interspecies transmission to the hamster.
Xu, Cheng; Guo, Tz-Chun; Mutoloki, Stephen; Haugland, Oyvind; Evensen, Oystein
Salmonid alphavirus subtype-3 (SAV-3) infection in Atlantic salmon is exclusively found in Norway. The salmonid alphaviruses have been well characterized at the genome level but there is limited information about the host-pathogen interaction phenomena. This study was undertaken to characterize the replication and spread of SAV-3 in internal organs of experimentally infected Atlantic salmon and the subsequent innate and adaptive immune responses. In addition, suitability of a cohabitation challenge model for this virus was also examined. Groups of fish were infected by intramuscular injection (IM), cohabited (CO) or kept uninfected in a separate tank. Samples of pancreas, kidney, spleen, heart and skeletal muscles were collected at 2, 4 and 8 weeks post infection (wpi). Pathological changes were assessed by histology concurrently with viral loads and mRNA expression of immune genes by real time RT-PCR. Pathological changes were only observed in the pancreas and heart (target organs) of both IM and CO groups, with changes appearing first in the pancreas (2 wpi) in the former. Lesions with increasing severity over time coincided with high viral loads despite significant induction of IFN-α, Mx and ISG15. IFN-γ and MHC-I were expressed in all tissues examined and their induction appeared in parallel with that of IL-10. Inflammatory genes TNF-α, IL-12 and IL-8 were only induced in the heart during pathology while T cell-related genes CD3ε, CD4, CD8, TCR-α and MHC-II were expressed in target organs at 8 wpi. These findings suggest that the onset of innate responses came too late to limit virus replication. Furthermore, SAV-3 infections in Atlantic salmon induce Th1/cytotoxic responses in common with other alphaviruses infecting higher vertebrates. Our findings demonstrate that SAV-3 can be transmitted via the water making it suitable for a cohabitation challenge model.
Broll, S; Alexandersen, S
The transplacental transmission of Aleutian mink disease parvovirus (ADV) was studied in experimental infection of 1-year-old female non-Aleutian mink. The ADV-seronegative female mink were inoculated with ADV prior to mating or after the expected implantation of the embryos during pregnancy. A group of uninfected females served as a control group. Animals from each group were killed prior to or shortly after parturition. The in situ hybridization technique with radiolabeled strand-specific RNA probes was used to determine target cells of virus infection and virus replication. In both infected groups, ADV crossed the endotheliochorial placental barrier, although animals infected before mating already had high antibody titers against ADV at the time of implantation. The percentage of dead and resorbed fetuses was much higher in dams infected before mating. In the placentae of these mink, virus DNA and viral mRNA were detected in cells in the mesenchymal stroma of the placental labyrinth and hematoma but only occasionally in the cytotrophoblast of the placental hematoma. Placentae of animals infected during pregnancy showed in addition very high levels of virus and also viral replication in a large number of cytotrophoblast cells in the placental hematoma, which exhibited distinct inclusion bodies. In both groups, neither virus nor virus replication could be detected in maternal endothelial cells or fetal syncytiotrophoblast of the placental labyrinth. Fetuses were positive for virus and viral replication at high levels in a wide range of tissues. Possible routes of transplacental transmission of ADV and the role of trophoblast cells as targets for viral replication are discussed. PMID:8627663
Lombard, Gian F.
At the first time, the CO2 laser was utilised in infective neurosurgical pathology as a surgical cutting instrument to remove inflammatory pseudomembranes in chronic osteomyelitis, and as a vaporising instmment on the dura mater surface. Successively, the instrument, defocused and at a low power, was used for prolonged and diffuse photo coagulation ofthe surgical cavity, particularly, ofthe dural surface and ofthe osteomyelitic bone edges, with the aim to sterilise tissues. So, we saw a shortening of the average time of wound healing and a lack of recurrence of the septic pathology. Then, we have treated, with CO2 laser, intracranial infective pathology: i.e. primary abscesses, capsulated or not, circumscribed purulent encephalitis, secondary abscesses in surgical cavities (patients operated for intracranial hematomas and tumors). In these cases we have obtained a lack of septic recurrences and an improvement ofneurological post-operative course. Thank to these results, we have continued to use laser in infective pathology; for giving an experimental support to these results we have carried on researches in vivo (on the experimental animal) to see the interaction between the laser and inflammatory tissue, and in vitro (on bacterial culture: in solid and liquid media) to see the laser effect on the bacterial cell. The bacterial cell has been also sensibiized to the photo dynamic effect of the laser (Argon, He-Ne), with hematoporphyrin. The goal of these experiments is to understand the role of thermal, photochemical, and mechanic resonance laser effects in the interaction between laser radiation and bacterial cell.
Vignoles, Philippe; Dreyfuss, Gilles; Rondelaud, Daniel
As large numbers of metacercariae of Fasciola hepatica are necessary for research, experimental infections of Galba truncatula and Pseudosuccinea columella with this digenean were carried out to determine the better intermediate host for metacercarial production and, consequently, the most profitable snail for decreasing the cost price of these larvae. Pre-adult snails (4 mm in shell height) originating from two populations per lymnaeid species were individually exposed to two or five miracidia, raised at 23 °C and followed for cercarial shedding up to their death. Compared to values noted in G. truncatula, the survival of P. columella on day 30 post-exposure was significantly greater, while the prevalence of F. hepatica infection was significantly lower. In the four P. columella groups, metacercarial production was significantly greater than that noted in the four groups of G. truncatula (347–453 per cercariae-shedding snail versus 163–275, respectively). Apart from one population of G. truncatula, the use of five miracidia per snail at exposure significantly increased the prevalence of F. hepatica in P. columella and the other population of G. truncatula, whereas it did not have any clear effect on the mean number of metacercariae. The use of P. columella for experimental infections with F. hepatica resulted in significantly higher metacercarial production than that noted with G. truncatula, in spite of a lower prevalence for the former lymnaeid. This finding allows for a significant decrease in the cost price of these larvae for commercial production. PMID:25907356
Vivas-Alegre, Santiago; Fernández-Natal, Isabel; López-Fidalgo, Eduardo; Rivero-Lezcano, Octavio Miguel
Experimental infections of either cells or animals require the preparation of good quality inocula. Unfortunately, the important pulmonary pathogen Streptococcus pneumoniae is a fastidious microorganism that suffers an autolysis process when cultured in vitro. Supplementation of Todd–Hewitt broth with a biological buffer (20 mM Tris–HCl, pH = 7.8) promotes a six hours delay in the beginning of the autolysis process. Additional improvements include washing bacteria before freezing, avoiding manipulations after thawing, and the use of glycerol (<18%) as a cryoprotectant, instead of reagents like skimmed milk that may affect cell cultures. With the proposed protocol >70% of the frozen bacteria was viable after 28 weeks at −80 °C, and aliquots were highly homogeneous. We have tested their utility in a whole blood infection model and have found that human plasma exhibits a higher microbicidal activity than whole blood, a result that we have not found previously reported. Additionally, we have also observed significant variations in the antimicrobial activity against different strains, which might be related to their virulence.•Media culture buffering extends S. pneumoniae viability for 6 h.•Washing before freezing of single use aliquots minimizes manipulation after thawing.•Experimental infection with the frozen inocula has shown that plasma has higher bactericidal activity than blood. PMID:26844211
Carrasco, Adriano de Oliveira Torres; Seki, Meire Christina; Benevenute, Jyan Lucas; Ikeda, Priscila; Pinto, Aramis Augusto
This study was designed with the goal of adding as much information as possible about the role of pigeons (Columba livia) and chickens (Gallus gallus) in Newcastle disease virus epidemiology. These species were submitted to direct experimental infection with Newcastle disease virus to evaluate interspecies transmission and virus-host relationships. The results obtained in four experimental models were analyzed by hemagglutination inhibition and reverse transcriptase polymerase chain reaction for detection of virus shedding. These techniques revealed that both avian species, when previously immunized with a low pathogenic Newcastle disease virus strain (LaSota), developed high antibody titers that significantly reduced virus shedding after infection with a highly pathogenic Newcastle disease virus strain (São Joao do Meriti) and that, in chickens, prevent clinical signs. Infected pigeons shed the pathogenic strain, which was not detected in sentinel chickens or control birds. When the presence of Newcastle disease virus was analyzed in tissue samples by RT-PCR, in both species, the virus was most frequently found in the spleen. The vaccination regimen can prevent clinical disease in chickens and reduce viral shedding by chickens or pigeons. Biosecurity measures associated with vaccination programs are crucial to maintain a virulent Newcastle disease virus-free status in industrial poultry in Brazil. PMID:26887250
Carrasco, Adriano de Oliveira Torres; Seki, Meire Christina; Benevenute, Jyan Lucas; Ikeda, Priscila; Pinto, Aramis Augusto
This study was designed with the goal of adding as much information as possible about the role of pigeons (Columba livia) and chickens (Gallus gallus) in Newcastle disease virus epidemiology. These species were submitted to direct experimental infection with Newcastle disease virus to evaluate interspecies transmission and virus-host relationships. The results obtained in four experimental models were analyzed by hemagglutination inhibition and reverse transcriptase polymerase chain reaction for detection of virus shedding. These techniques revealed that both avian species, when previously immunized with a low pathogenic Newcastle disease virus strain (LaSota), developed high antibody titers that significantly reduced virus shedding after infection with a highly pathogenic Newcastle disease virus strain (São Joao do Meriti) and that, in chickens, prevent clinical signs. Infected pigeons shed the pathogenic strain, which was not detected in sentinel chickens or control birds. When the presence of Newcastle disease virus was analyzed in tissue samples by RT-PCR, in both species, the virus was most frequently found in the spleen. The vaccination regimen can prevent clinical disease in chickens and reduce viral shedding by chickens or pigeons. Biosecurity measures associated with vaccination programs are crucial to maintain a virulent Newcastle disease virus-free status in industrial poultry in Brazil.
Vignoles, Philippe; Dreyfuss, Gilles; Rondelaud, Daniel
As large numbers of metacercariae of Fasciola hepatica are necessary for research, experimental infections of Galba truncatula and Pseudosuccinea columella with this digenean were carried out to determine the better intermediate host for metacercarial production and, consequently, the most profitable snail for decreasing the cost price of these larvae. Pre-adult snails (4 mm in shell height) originating from two populations per lymnaeid species were individually exposed to two or five miracidia, raised at 23 °C and followed for cercarial shedding up to their death. Compared to values noted in G. truncatula, the survival of P. columella on day 30 post-exposure was significantly greater, while the prevalence of F. hepatica infection was significantly lower. In the four P. columella groups, metacercarial production was significantly greater than that noted in the four groups of G. truncatula (347-453 per cercariae-shedding snail versus 163-275, respectively). Apart from one population of G. truncatula, the use of five miracidia per snail at exposure significantly increased the prevalence of F. hepatica in P. columella and the other population of G. truncatula, whereas it did not have any clear effect on the mean number of metacercariae. The use of P. columella for experimental infections with F. hepatica resulted in significantly higher metacercarial production than that noted with G. truncatula, in spite of a lower prevalence for the former lymnaeid. This finding allows for a significant decrease in the cost price of these larvae for commercial production.
Carneiro, Liliane Almeida; Silveira, Fernando Tobias; Campos, Marliane Batista; Brígido, Maria do Carmo de Oliveira; Gomes, Claudia Maria C; Corbett, Carlos E P; Laurenti, Márcia D
In Amazonian Brazil, the Cebus apella monkey (Primates: Cebidae) has been associated with the enzootic cycle of Leishmania (V.) shawi, a dermotropic parasite causing American cutaneous leishmaniasis (ACL). It has also been successfully used as animal model for studying cutaneous leishmaniasis. In this work, there has been investigated its susceptibility to experimental Leishmania (L.) infantum chagasi-infection, the etiologic agent of American visceral leishmaniasis (AVL). There were used ten C. apella specimens, eight adult and two young, four males and six females, all born and raised in captivity. Two experimental infection protocols were performed: i) six monkeys were inoculated, intra-dermal via (ID), into the base of the tail with 2 x 10(6) promastigotes forms from the stationary phase culture medium; ii) other four monkeys were inoculated with 3 x 10(7) amastigotes forms from the visceral infection of infected hamsters by two different via: a) two by intravenous via (IV) and, b) other two by intra-peritoneal via (IP). The parameters of infection evaluation included: a) clinical: physical exam of abdomen, weigh and body temperature; b) parasitological: needle aspiration of the bone-marrow for searching of amastigotes (Giemsa-stained smears) and promastigotes forms (culture medium); c) immunological: Indirect fluorescence antibody test (IFAT) and, Delayed-type hypersensitivity (DTH). In the six monkeys ID inoculated (promastigotes forms) all parameters of infection evaluation were negative during the 12 months period of follow-up. Among the four monkeys inoculated with amastigotes forms, two IV inoculated showed the parasite in the bone-marrow from the first toward to the sixth month p.i. and following that they cleared the infection, whereas the other two IP inoculated were totally negative. These four monkeys showed specific IgG-antibody response since the third month p.i. (IP: 1/80 and IV: 1/320 IgG) toward to the 12th month (IP: 1/160 and IV: 1/5120). The
Fentie, Tsegaw; Dadi, Kara; Kassa, Tesfu; Sahle, Mesfin; Cattoli, Giovanni
An experimental study was conducted to evaluate the effect of vaccines produced in Ethiopia from vaccine strains used worldwide on the transmission characteristics of velogenic Newcastle disease virus field strain after different vaccination schemes. Chickens were vaccinated with Hitchner B1, La Sota or I-2 via the intraocular and intranasal routes. Vaccine and challenge viruses induced high antibody levels, both in inoculated and contact birds. Prime-boost vaccination protected birds against morbidity and mortality and significantly reduced the incidence of viral shedding from chickens compared with single vaccinated and unvaccinated birds. Protection from disease and mortality was correlated with the presence of positive antibody titres (>4 log2) at day of challenge. Most of the unvaccinated and in-contact birds excreted the virus and showed a high level of antibody titres, indicating the high infectivity of the challenge virus. The detection of the challenge virus in most of vaccinated birds demonstrated that the tested vaccination protocols cannot fully protect birds from viral infection, replication and shedding, and vaccinated-infected birds can act as a source of infection for susceptible flocks. The high mortality observed in unvaccinated birds and their contacts confirmed the virulence of the challenge virus and indicated that this field virus strain can easily spread in an unvaccinated poultry population and cause major outbreaks. Progressive vaccinations supported by biosecurity measures should therefore be implemented to control the disease and introduction of the virus to the poultry farms.
Xu, Yin; Tian, Chan; Wang, Shao-Bin; Xie, Wu-Ling; Guo, Yan; Zhang, Jin; Shi, Qi; Chen, Cao; Dong, Xiao-Ping
Macroautophagy is an important process for removing misfolded and aggregated protein in cells, the dysfunction of which has been directly linked to an increasing number of neurodegenerative disorders. However, the details of macroautophagy in prion diseases remain obscure. Here we demonstrated that in the terminal stages of scrapie strain 263K-infected hamsters and human genetic prion diseases, the microtubule-associated protein 1 light chain 3 (LC3) was converted from the cytosolic form to the autophagosome-bound membrane form. Macroautophagy substrate sequestosome 1 (SQSTM1) and polyubiquitinated proteins were downregulated in the brains of sick individuals, indicating enhanced macroautophagic protein degradation. The levels of mechanistic target of rapamycin (MTOR) and phosphorylated MTOR (p-MTOR) were significantly decreased, which implies that this enhancement of the macroautophagic response is likely through the MTOR pathway which is a negative regulator for the initiation of macroautophagy. Dynamic assays of the autophagic system in the brains of scrapie experimental hamsters after inoculation showed that alterations of the autophagic system appeared along with the deposits of PrPSc in the infected brains. Immunofluorescent assays revealed specific staining of autophagosomes in neurons that were not colocalized with deposits of PrPSc in the brains of scrapie infected hamsters, however, autophagosome did colocalize with PrPSc in a prion-infected cell line after treatment with bafilomycin A1. These results suggest that activation of macroautophagy in brains is a disease-correlative phenomenon in prion diseases. PMID:22874564
Boada-Sucre, Alpidio A.; Rossi Spadafora, Marcello Salvatore; Tavares-Marques, Lucinda M.; Finol, Héctor J.; Reyna-Bello, Armando
Trypanosomosis, a globally occurring parasitic disease, poses as a major obstacle to livestock production in tropical and subtropical regions resulting in tangible economic losses. In Latin America including Venezuela, trypanosomosis of ruminants is mainly caused by Trypanosoma vivax. Biologically active substances produced from trypanosomes, as well as host-trypanosome cellular interactions, contribute to the pathogenesis of anemia in an infection. The aim of this study was to examine with a scanning electron microscope the cellular interactions and alterations in ovine red blood cells (RBC) experimentally infected with T. vivax. Ovine infection resulted in changes of RBC shape as well as the formation of surface holes or vesicles. A frequent observation was the adhesion to the ovine RBC by the trypanosome's free flagellum, cell body, or attached flagellum in a process mediated by the filopodia emission from the trypanosome surface. The observed RBC alterations are caused by mechanical and biochemical damage from host-parasite interactions occurring in the bloodstream. The altered erythrocytes are prone to mononuclear phagocytic removal contributing to the hematocrit decrease during infection. PMID:27293960
DeVincenzo, John P.; Wilkinson, Tom; Vaishnaw, Akshay; Cehelsky, Jeff; Meyers, Rachel; Nochur, Saraswathy; Harrison, Lisa; Meeking, Patricia; Mann, Alex; Moane, Elizabeth; Oxford, John; Pareek, Rajat; Moore, Ryves; Walsh, Ed; Studholme, Robert; Dorsett, Preston; Alvarez, Rene; Lambkin-Williams, Robert
Rationale: Respiratory syncytial virus (RSV) is the leading cause of childhood lower respiratory infection, yet viable therapies are lacking. Two major challenges have stalled antiviral development: ethical difficulties in performing pediatric proof-of-concept studies and the prevailing concept that the disease is immune-mediated rather than being driven by viral load. Objectives: The development of a human experimental wild-type RSV infection model to address these challenges. Methods: Healthy volunteers (n = 35), in five cohorts, received increasing quantities (3.0–5.4 log plaque-forming units/person) of wild-type RSV-A intranasally. Measurements and Main Results: Overall, 77% of volunteers consistently shed virus. Infection rate, viral loads, disease severity, and safety were similar between cohorts and were unrelated to quantity of RSV received. Symptoms began near the time of initial viral detection, peaked in severity near when viral load peaked, and subsided as viral loads (measured by real-time polymerase chain reaction) slowly declined. Viral loads correlated significantly with intranasal proinflammatory cytokine concentrations (IL-6 and IL-8). Increased viral load correlated consistently with increases in multiple different disease measurements (symptoms, physical examination, and amount of nasal mucus). Conclusions: Viral load appears to drive disease manifestations in humans with RSV infection. The observed parallel viral and disease kinetics support a potential clinical benefit of RSV antivirals. This reproducible model facilitates the development of future RSV therapeutics. PMID:20622030
Avila-Villa, Luz A.; Gollas-Galván, Teresa; Martínez-Porchas, Marcel; Mendoza-Cano, Fernando; Hernández-López, Jorge
Necrotizing hepatopancreatitis bacterium (NHPB) is an obligated intracellular bacteria causing severe hepatopancreatic damages and mass mortalities in penaeid shrimp. The worldwide distribution of penaeid shrimp as alien species threatens the life cycle of other crustacean species. The aim of the experiment was to evaluate the possibility of experimentally infecting the American lobster (Homarus americanus) with NHPB extracted from shrimp hepatopancreas. Homogenates from infected shrimp were fed by force to lobsters. Other group of lobsters was fed with homogenates of NHPB-free hepatopancreas. After the 15th day from initial inoculation, the presence of NHPB was detected by polymerase chain reaction in feces and hepatopancreas from lobsters inoculated with infected homogenates. Necrotized spots were observed in the surface of lobster hepatopancreas. In contrast, lobsters fed on NHPB-free homogenates resulted negative for NHPB. Evidence suggests the plasticity of NHPB which can infect crustacean from different species and inhabiting diverse latitudes. Considering the results, the American lobster could be a good candidate to maintain available NHPB in vivo. PMID:22645497
Tollenaere, C; Rahalison, L; Ranjalahy, M; Duplantier, J-M; Rahelinirina, S; Telfer, S; Brouat, C
In Madagascar, the black rat, Rattus rattus, is the main reservoir of plague (Yersinia pestis infection), a disease still responsible for hundreds of cases each year in this country. This study used experimental plague challenge to assess susceptibility in wild-caught rats to better understand how R. rattus can act as a plague reservoir. An important difference in plague resistance between rat populations from the plague focus (central highlands) and those from the plague-free zone (low altitude area) was confirmed to be a widespread phenomenon. In rats from the plague focus, we observed that sex influenced plague susceptibility, with males slightly more resistant than females. Other individual factors investigated (weight and habitat of sampling) did not affect plague resistance. When infected at high bacterial dose (more than 10⁵ bacteria injected), rats from the plague focus died mainly within 3-5 days and produced specific antibodies, whereas after low-dose infection (< 5,000 bacteria), delayed mortality was observed and surviving seronegative rats were not uncommon. These results concerning plague resistance level and the course of infection in the black rat would contribute to a better understanding of plague circulation in Madagascar.
Ikeda, Y; Fukuoka, Y; Motomura, K; Yasuda, T; Nishino, T
In previous papers (Y. Ikeda and T. Nishino, Antimicrob. Agents Chemother. 32:1073-1077, 1988; Y. Ikeda, T. Nishino, and T. Tanino, Antimicrob. Agents Chemother. 31:865-869, 1987), we reported that many of the 7-aminothiazolyl cephalosporins, such as cefmenoxime, showed paradoxically reduced activity against Proteus vulgaris at higher concentrations, whereas these paradoxical effects were not observed for other types of cephalosporins, such as cefbuperazone and cefoperazone. In this study, we compare the therapeutic effect of cefmenoxime with that of cefbuperazone and explore the in vivo paradoxical effect of cefmenoxime by using an experimental infection model in mice. In an intraperitoneal infection with P. vulgaris 11, the survival rate with cefmenoxime was increased to 43% at 3.13 mg/kg but was lower at higher doses. On the other hand, cefbuperazone did not show such a paradoxical therapeutic effect. In mice infected with P. vulgaris 11, cefmenoxime levels in both serum and peritoneal washings were rapidly reduced and beta-lactamase activities in the peritoneal cavity were increased at higher cefmenoxime doses. These findings suggested that high levels of cefmenoxime at the infection site induced increased production of beta-lactamase, which then rapidly inactivated the antibiotic. We conclude that the paradoxical therapeutic effect of cefmenoxime against P. vulgaris occurs by the same mechanisms as the in vitro effect and that the high beta-lactamase inducibility and low beta-lactamase stability may account for the paradoxical therapeutic effect of cefmenoxime against P. vulgaris.
Castrucci, G; Osburn, B I; Ferrari, M; Traldi, V
This presentation summarizes the results of a study on the pathogenesis of bovine viral diarrhea (BVDV) infection. The cytopathic (CP) strain TVM-2 of BVDV induced in calves an overt clinical disease which is usually recorded as the acute primary BVDV infection observed under natural conditions. In contrast the non-cytopathic (NCP) strain New York-1 of BVDV did not cause any significant signs of disease. However, when the calves were immunosuppressed by treatment with dexamethasone (DMS) the biotype of BVDV involved did not seem to be as important as it appeared to be in an immunologically normal animal. This was shown in this study by the NCP BVDV which caused a fatal disease in calves treated with DMS. A mixed infection given to calves by injecting them with both CP and NCP BVDV, did not result in any particularly serious disease. So, the potential immunosuppressive activity of BVDV itself for the host has not been proven under the experimental procedures used in this experiment. Finally, a modified-live CP BVDV vaccine was unable to cause clinical disease when injected into calves that had been infected previously with strain New York-1 of BVDV.
Lee, Hyeyeoun; Jeong, Hansol; Park, Surim; Yang, Myeon-Sik; Kim, Jongwon; Bae, Jaehyun; Kwon, Yonghwan; Kim, Min-Su; Oem, Jae-Ku; Lee, Myoung-Heon; Lim, Chae-Woong; Kim, Bumseok
Akabane virus (AKAV), an arthropod-transmitted bunyavirus, is a major cause of congenital abnormalities and encephalomyelitis in ruminants. In 2010, there was a major outbreak of encephalomyelitis in Korea and fifteen AKAV strains, including AKAV-7, were isolated from cows. To identify the neuropathogenicity of AKAV-7, we performed experimental infection of cows. Six-month-old female Korean Holstein dairy cattle were inoculated with AKAV-7 by various routes, including intracerebral (IC), intrasubarachnoid space (IS), subcutaneous (SC) and intravenous (IV); a separate group was vaccinated before intravenous infection. Five of the six cows in the IC group and two of the six cows in the IS group showed clinical signs such as locomotor ataxia and paralysis of the hind limbs. Three of six cows died after IC infection 9-12 days post infection (dpi). Histopathologic changes such as nonsuppurative encephalomyelitis were confirmed in various parts of the central nervous system in the IC, IS and SC groups. Early onset of neutralizing antibodies in the serum and lower viral mRNA levels in the peripheral blood mononuclear cells (PBMCs) and various tissues in the vaccinated group was noticeable compared to the unvaccinated group (IV group). We suggest that the AKAV vaccine currently used in Korea may be partially effective for protection against AKAV-7 in cows.
Brazão, Vânia; Del Vecchio Filipin, Marina; Santello, Fabricia Helena; Caetano, Leony Cristina; Abrahão, Ana Amélia Carraro; Toldo, Míriam Paula Alonso; do Prado, José Clóvis
Melatonin by exhibiting antioxidant, anti-aging, and immunomodulatory properties favorably modulate the immune function, protecting the hosts from several infectious diseases. Zinc is an essential trace element important for the efficiency of the immune system in reason of its widespread role in the activity of enzymes, transcription factors and cytokines. The etiology of Chagas' disease, caused by a protozoan parasite Trypanosoma cruzi, has been the focus of considerable discussion, although chronic phase still remains not fully understood. This study showed that zinc and melatonin treatment did not affect the percentage of both CD4+ and CD8+ T lymphocytes subsets in chronically infected animals. Increased levels of IL-2 and IL-10, as well as, enhanced thymocyte proliferation in T. cruzi infected groups under zinc and melatonin therapy was observed as compared to untreated group. Conversely, during the chronic phase of infection, macrophages counts were reduced in melatonin and zinc-melatonin treated animals. The combined actions of zinc and melatonin have beneficial effects in counteracting parasite-induced immune dysregulation, protecting animals against the harmful actions of chronic T. cruzi infection. Furthermore, our results provide an experimental basis for further studies on the role of immunomodulatory therapies.
Berthelot-Hérault, F; Cariolet, R; Labbé, A; Gottschalk, M; Cardinal, J Y; Kobisch, M
A standardized model of Streptococcus suis type 2 infection in specific-pathogen-free piglets, housed in high-security barns, was used to compare the virulence of 3 French field strains of S. suis serotype 2 isolated from tonsils of a healthy pig (strain 65) or from diseased pigs (meningitis, strain 166', or septicemia, strain 24). In one of the 2 trials, 7-week-old pigs, in 3 groups of 8, were inoculated intravenously with 2 x 10(8) colony-forming units of S. suis type 2. In each group, 1 uninfected animal was a sentinel. Eight animals were also used as negative control group. The experiment was repeated under similar conditions with strains 65 and 166'. Virulence differed markedly among these S. suis strains when clinical signs, zootechnical performances, lesions, and bacteriological data were analyzed. Strain 65 did not induce clinical signs in inoculated pigs. In contrast, pigs infected with the other 2 strains exhibited clinical signs and typical lesions of S. suis type 2 infections. Differences in virulence were also observed between the 2 virulent strains. Sentinel animals exhibited the same manifestations as those recorded in inoculated piglets. Results were similar in the second trial, indicating that under the present experimental conditions, results were reproducible. The standardized conditions described in this study could be a useful tool to further study about the S. suis infection. PMID:11480526
Inceboz, Tonay; Lambrecht, Fatma Yurt; Eren, Mine Şencan; Girginkardeşler, Nogay; Bekiş, Recep; Yilmaz, Osman; Er, Özge; Özbilgin, Ahmet
We aimed to assess the ability of (131)I-Pentamidine scintigraphy to detect the lesions of Leishmania tropica infection. An experimental model of cutaneous leishmaniasis was developed. The presence of cutaneous leishmaniasis was confirmed. Pentamidine was radioiodinated with (131)I. The radiolabeled pentamidine was validated by the requisite quality control tests to check its radiolabeling efficiency, in vitro stability. (131)I-Pentamidine (activity: 18.5 MBq/100 µl) was injected intracardiacally into infected hamsters. Static whole body images of the hamsters were acquired under the gamma camera at 5 and 30 min, 2, 6 and 24 h following the administration. On the scintigrams, anatomically adjusted regions of interest (ROIs) were drawn over the right feet (target) and left feet (not-target) and various organs. Accumulation of (131)I-Pentamidine at sites of infection is expressed as the target to non-target (T/NT) ratio. The results T/NT ratio decreased with time. In concluding the (131)I-Pentamidine has poor sensitivity in detection of L. tropica infection.
Marques, P. X.; Souda, Puneet; O'Donovan, J.; Gutierrez, J.; Gutierrez, E. J.; Worrall, S.; McElroy, M.; Proctor, A.; Brady, C.; Sammin, D.; Basset, H. F.; Whitelegge, Julian P.; Markey, B. E.; Nally, J. E.
Chlamydophila abortus is an intracellular pathogen and the etiological agent of enzootic abortion of ewes (EAE). C. abortus has a biphasic development cycle; extracellular infectious elementary bodies (EB) attach and penetrate host cells, where they give rise to intracellular, metabolically active reticulate bodies (RB). RB divide by binary fission and subsequently mature to EB, which, on rupture of infected cells, are released to infect new host cells. Pregnant ewes were challenged with 2 × 106 inclusion forming units (IFU) of C. abortus cultured in yolk sac (comprising both EB and RB). Serum samples were collected at 0, 7, 14, 21, 27, 30, 35, 40, and 43 days postinfection (dpi) and used to identify antigens of C. abortus expressed during disease. Additionally, sera from fetal lambs were collected at 30, 35, 40, and 43 dpi. All serum samples collected from experimentally infected pregnant ewes reacted specifically with several antigens of EB as determined by one-dimensional (1-D) and 2-D gel electrophoresis; reactive antigens identified by mass spectrometry included the major outer membrane protein (MOMP), polymorphic outer membrane protein (POMP), and macrophage infectivity potentiator (MIP) lipoprotein. PMID:20554807
Grilli, E; Tugnoli, B; Formigoni, A; Massi, P; Fantinati, P; Tosi, G; Piva, A
The reduction of Salmonella prevalence in broilers is a priority in European Union agricultural policies because treatment with antibiotics is forbidden by Regulation (EC) 2160/2003. Two trials were conducted to evaluate the efficacy of a microencapsulated blend of sorbic acid and nature-identical compounds (i.e., chemically synthesized botanicals; SAB) on the reduction of the cecal prevalence and contents of Salmonella enterica serovars Hadar and Enteritidis in experimentally infected chickens. In the first trial, 125 one-day-old Lohmann specific-pathogen-free chickens were assigned to one of the following treatments: negative control (not challenged and not treated), positive control (challenged and not treated), SAB0.3, SAB1, or SAB5 (challenged and treated with the microencapsulated blend included in the feed at 0.03, 0.1, or 0.5%, respectively). At 30 d of age, birds were infected with 10(6) cfu of Salmonella Hadar, and after 5, 10, or 20 d postinfection, 5, 10, and 10 birds per treatment, respectively, were killed and the cecal contents and liver and spleen samples were analyzed for Salmonella Hadar. In the second trial, 100 one-day-old Ross 708 chickens were assigned to 1 of 5 treatments: control (not treated), SAB0.3, SAB1, SAB2, or SAB5 (treated with the blend included in the feed at 0.03, 0.1, 0.2, or 0.5%, respectively). At 7 d of age, the birds were challenged with 10(5) cfu of Salmonella Enteritidis, and after 7, 14, or 24 d after challenge, 5, 5, and 10 birds per treatment, respectively, were killed and cecal contents were analyzed for Salmonella Enteritidis. Results showed that in the early stage of infection Salmonella prevalence was high in both studies, whereas at the end of the observation periods, the blends at 0.03, 0.1, and 0.5 in the challenge with Salmonella Hadar and at 0.2 and 0.5% in the challenge with Salmonella Enteritidis significantly reduced (by 2 log(10) cfu) the cecal content of Salmonella. This study showed that intestinal
Chryssafidis, Andreas L; Cantón, Germán; Chianini, Francesca; Innes, Elisabeth A; Madureira, Ed H; Soares, Rodrigo M; Gennari, Solange M
The water buffalo (Bubalus bubalis) is an important species in several countries for its milk and meat production, as well as for transport and other agricultural activities. It is, in general, considered more resistant than cattle to different parasitic diseases, also less demanding for forage quality. It has been postulated that buffalo may be resistant to abortion caused by neosporosis, because of high serological prevalences found in buffalo herds from different localities, with no description of Neospora caninum-related abortion. Recent studies have demonstrated the potential impact of neosporosis in pregnant water buffalo cows. In this work, three pregnant buffalo cows were experimentally infected with Nc-1 strain of N. caninum, and abortion was detected 35 days post-infection. Molecular and histopathological results found in post-mortem tissues are described and discussed, confirming the susceptibility of water buffalos to abortion caused by N. caninum.
Deardorff, Eleanor R.; Forrester, Naomi L.; Travassos da Rosa, Amelia P.; Estrada-Franco, Jose G.; Navarro-Lopez, Roberto; Tesh, Robert B.
In 1993, an outbreak of encephalitis among 125 affected equids in coastal Chiapas, Mexico, resulted in a 50% case-fatality rate. The outbreak was attributed to Venezuelan equine encephalitis virus (VEEV) subtype IE, not previously associated with equine disease and death. To better understand the ecology of this VEEV strain in Chiapas, we experimentally infected 5 species of wild rodents and evaluated their competence as reservoir and amplifying hosts. Rodents from 1 species (Baiomys musculus) showed signs of disease and died by day 8 postinoculation. Rodents from the 4 other species (Liomys salvini, Oligoryzomys fulvescens, Oryzomys couesi, and Sigmodon hispidus) became viremic but survived and developed neutralizing antibodies, indicating that multiple species may contribute to VEEV maintenance. By infecting numerous rodent species and producing adequate viremia, VEEV may increase its chances of long-term persistence in nature and could increase risk for establishment in disease-endemic areas and amplification outside the disease-endemic range. PMID:19331726
Bandini, Luciana A; Neto, Aldo F A; Pena, Hilda F J; Cavalcante, Guacyara T; Schares, Gereon; Nishi, Sandra M; Gennari, Solange M
Neospora caninum is widely distributed in the world and this parasite is one of the major causes of abortion in cattle. Dogs and coyotes are definitive hosts of N. caninum and several species of domestic and wild animals are intermediate hosts. Dogs can become infected by the ingestion of tissues containing cysts and then excrete oocysts. It is not yet known whether sporulated oocysts are able to induce a patent infection in dogs, i.e. a shedding of N. caninum oocysts in feces. The objective of this study was to experimentally examine the infection of dogs by sporulated oocysts. The oocysts used in the experiment were obtained by feeding dogs with brain of buffaloes (Bubalus bubalis) positive for anti-N. caninum antibodies by indirect fluorescent antibody test (IFAT ≥200). Oocysts shed by these dogs were confirmed to be N. caninum by molecular methods and by bioassay in gerbils, and sporulated N. caninum oocysts were used for the oral infection of four dogs. The dogs were 8 weeks old and negative for antibodies to N. caninum and Toxoplasma gondii. Dogs 1 and 4 received an inoculum of 10,000 sporulated oocysts each; dog 2 an inoculum of 5000 sporulated oocysts and dog 3 received 1000 sporulated oocysts of N. caninum. The total feces excreted by these dogs were collected and examined daily for a period of 30 days. No oocysts were found in their feces. The dogs were monitored monthly for a 6-month period to observe a possible seroconversion and when this occurred the animals were eliminated from the experiment. Dogs 1 and 4 seroconverted 1 month after the infection with titer, in the IFAT, of 1600 and 800, respectively; the other two dogs presented no seroconvertion during the 6-month period. Dogs 1 and 2 were euthanized 180 days after infection and were examined for the detection of N. caninum in tissues (brain, muscle, lymph node, liver, lung, heart and bone marrow) by immunohistochemistry and PCR with negative results in both techniques. Bioassay in gerbils with
Laban, Linet T; Anjili, Christopher O; Mutiso, Joshua M; Ingonga, Johnstone; Kiige, Samuel G; Ngedzo, Mgala M; Gicheru, Michael M
Objective(s): Solanum acueastrum Dunal. has been shown to have some chemotherapeutic value. Leaf and berry water and methanol compounds of S. acueastrum were evaluated for possible antileishmanial activity In vivo on BALB/c mice and in vitro against Leishmania major promastigotes, amastigotes and vero cells. Materials and Methods: Dry S. aculeastrum berry and leaf material were extracted in methanol and water. L. major parasites were exposed to different concentrations of S. aculeastrum fruit and leaf compounds and the IC50 on the promastigotes, percentage of infection rate of macrophages by amastigotes and the toxicological effect on vero cells were determined. BALB/c mice were infected subcutaneously with 1×106 promastigotes and kept for four weeks to allow for disease establishment. Infected mice were treated with fruit and leaf methanolic and water compounds, amphotericin B (AmB), and sterile phosphate buffered saline (PBS). Results: Fruit methanol compound was most effective in inhibiting the growth of promastigotes with IC5078.62 μg/ml. Fruit water compound showed the best activity in inhibiting infection of macrophages by amastigotes. Fruit methanol compound was more toxic at Ld50=8.06 mg/ml to vero cells than amphotericin B. Analysis of variance computation indicated statistically significant difference in lesion sizes between experimental and control mice groups (P=0.0001). Splenic impression smears ANOVA indicated a highly significant difference in parasitic numbers between the experimental and the control groups (P=0.0001). Conclusion: The results demonstrate that compounds from S. aculeastrum have potential anti-leishmanial activities and the medicinal use of the plant poses considerable toxicity against dividing vero cells. PMID:25810878
Bambou, J C; Archimède, H; Arquet, R; Mahieu, M; Alexandre, G; González-Garcia, E; Mandonnet, N
The aim of the present study was to test the effect of dietary supplementation on resistance to experimental infection with Haemonchus contortus in Creole kids. One trial with three replicates involved a total of 154 female kids that were chosen from three successive cohorts of the Creole flock of INRA-Gardel in 2007. The kids were placed into four treatments according to the amount of concentrate they received: G0 (no concentrate and a quality Dichantium spp. hay ad libitum, HAY), G1 (HAY+100g commercial concentrate d(-1)), G2 (HAY+200 g commercial concentrate d(-1)), G3 (HAY+300 g commercial concentrate d(-1)). The G0-G3 groups were infected with a single dose of 10,000 H. contortus third stage larvae (L(3)) at Day 0 (D0). Each infected group was comprised of one half resistant and one half susceptible genetically indexed kids. The average breeding values on egg excretion at 11 months of age were distant of 0.70, 0.65, 0.61 and 0.61 genetic standard deviations in G0, G1, G2 and G3, respectively. The faecal egg count (FEC), packed cell volume (PCV), eosinophilia (EOSI) and dry matter intake (DMI) indices were monitored weekly until 42 days post-infection. Enzyme-linked immunosorbent assay was carried out on serum samples to determine the level of IgA anti-H. contortus L(3) crude extracts and adult excretion/secretion products (ESP). The 10,000 L(3) dose received by the kids induced a severe infection: 8000 eggs per gram at the FEC peak, a PCV less than 15% and mortality. Interestingly, the supplemented animals in G3 showed a higher level of EOSI but a lower level of IgA anti-L3 and IgA anti-ESP than non-supplemented animals (G0). Resistant and susceptible kids had significantly different FEC variations within the groups. Susceptible kids had a 1.6 times higher egg output than resistant kids in G0. This difference was not found in the supplemented groups. The results of this study showed that supplementary feeding improved resistance of Creole kids to H. contortus
The objective of the current research was to determine if exogenous melatonin would exert a “protective” effect on the gastrointestinal tract of sheep and prevent or reduce the horizontal transfer of E. coli O157:H7 from experimentally-infected to non-infected or “naïve” sheep. Sixteen crossbred ewe...
A comparative study was conducted using data from naive bison (n=45) and cattle (n=46) from 8 and 6 studies, respectively, in which a standardized Brucella abortus strain 2308 experimental challenge was administered. The incidence of abortion, fetal infection, uterine or mammary infection, or infec...
The objective of this paper is to study shedding patterns of cows infected with Mycobacterium avium subsp. paratuberculosis (MAP). While multiple single farm studies of MAP dynamics were reported, there is not large-scale meta-analysis of both natural and experimental infections. Large difference...
Reppert, E; Galindo, R C; Breshears, M A; Kocan, K M; Blouin, E F; de la Fuente, J
Anaplasma phagocytophilum, first identified as a pathogen of sheep in Europe, more recently has been recognized as an emerging tick-borne pathogen of humans in the U.S. and Europe. Transmission of A. phagocytophilum is reported to be by ticks, primarily of the genus Ixodes. While mechanical and transplacental transmission of the type genus organism, A. marginale, occur in addition to tick transmission, these modes of transmission have not been considered for A. phagocytophilum. Recently, we developed a sheep model for studying host-tick-pathogen interactions of the human NY-18 A. phagocytophilum isolate. Sheep were susceptible to infection with this human isolate and served as a source of infection for I. scapularis ticks, but they did not display clinical signs of disease, and the pathogen was not apparent in stained blood smears. In the course of these experiments, one sheep unexpectedly gave birth to a lamb 5 weeks after being experimentally infected by inoculation with the pathogen propagated in HL-60 cells. The lamb was depressed and not feeding and was subsequently euthanized 18 h after birth. Tissues were collected at necropsy for microscopic examination and PCR to confirm A. phagocytophilum infection. At necropsy, the stomach contained colostrum, the spleen was moderately enlarged and thickened with conspicuous lymphoid follicles, and mesenteric lymph nodes were mildly enlarged and contained moderate infiltrates of eosinophils and neutrophils. Blood, spleen, heart, skin and cervical and mesenteric lymph nodes tested positive for A. phagocytophilum by PCR, and sequence analysis confirmed that the lamb was infected with the NY-18 isolate. Transplacental transmission should therefore be considered as a means of A. phagocytophilum transmission and may likely contribute to the epidemiology of tick-borne fever in sheep and other mammals, including humans.
Afzali, Seyedeh F.; Mohd Daud, Hassan Hj.; Sharifpour, Issa; Afsharnasab, Mohammad; Shankar, Shiv
Aim: Epizootic ulcerative syndrome (EUS) causes by aquatic oomycete fungus, Aphanomyces invadans is a dangerous fish disease of a wide range of fresh and brackish water, wild and farmed fish throughout the world. The objective of the present study was to determine the susceptibility of a number of tropical fish species to the EUS and compare the severity of infection between experimental groups. Materials and Methods: Snakehead, Channa striata (Bloch, 1793); snakeskin gourami, Trichopodus pectoralis (Regan, 1910); koi carp, Cyprinus carpio (Linnaeus, 1758); broadhead catfish, Clarias macrocephalus (Günther, 1864); goldfish, Carassius auratus (Linnaeus, 1758); climbing perch, Anabas testudineus (Bloch, 1792); and Nile tilapia, Oreochromis niloticus (Linnaeus, 1758) were challenged by intramuscular injection using zoospores of Aphanomyces invadans (NJM9701). The infected fish skins and muscles were examined for EUS histopathological characteristics, and the results on the severity of lesions and mortality were analyzed using SPSS program. Results: All zoospore-injected fish were shown to be susceptible to the EUS infection except Nile tilapia. Although, the general histopathological pattern was similar in the zoospore-injected group, but there were some variation in granulomatous reaction, that is the presence or absence of giant cells, and time of mortality were detected. The result of statistical analysis showed that there was a significant difference between species, (c2=145.11 and p<0.01). Conclusion: Gourami, koi carp, and catfish were demonstrated to be highly susceptible while goldfish and climbing perch were found to be moderately susceptible to the EUS infection. These findings suggested that the cellular response of fish to mycotic infection and granulomatous reaction varied in different fish species, which could not be an indicator of susceptibility or resistant to the EUS itself, although it was shown that the granulation rate and the level of maturity
Newbould, F. H. S.
Experimental infections were produced in 78 quarters of 17 cows by the infusion of small numbers of a single strain of Staphylococcus aureus. In each single experiment three quarters in a cow were infected, with the fourth left as a control. At times varying from three to 60 days after the infusion of organisms, a standard intramammary antibiotic treatment was administered on a single occasion. A cure was arbitrarily defined as the absence of the organism in foremilk, from direct plating and replated incubated milk, together with return to normal somatic cell count levels as determined by an electronic counter. With these standardized conditions the effects of a number of cow associated factors on the outcome of the therapy were determined. Forty-three of the 78 quarters (55%) were cured by the standard treatment. Significant differences in percentages of quarters cured were found to be associated with the duration of infection before therapy, the lactation age of the cow, the length of time in lactation, somatic cell count in milk at time of treatment, the location of the quarter in the udder and individual cows. No significant effects on the outcome of the standard treatment were found associated with the number of bacteria in the secretion at the time of treatment, previous infection and cure in a quarter nor the season of the year in which treatment was given. Of the 35 quarters in which infection recurred following treatment, organisms were reisolated from 12 within four days, 18 between five and nine days, four between ten and 17 days and one after 28 days. From these data it is apparent that if, as has been suggested, models such as described are to be used for efficacy trials, standardization of some parameters is essential. PMID:4279760
Neumann, Arthur da Silva; Dias, Felipe de Almeida; Ferreira, Jéssica da Silva; Fontes, Amanda Nogueira Brum; Rosa, Patricia Sammarco; Macedo, Rafael Enrique; Oliveira, José Henrique; Teixeira, Raquel Lima de Figueiredo; Pessolani, Maria Cristina Vidal; Moraes, Milton Ozório; Suffys, Philip Noel; Oliveira, Pedro L.; Sorgine, Marcos Henrique Ferreira; Lara, Flavio Alves
Leprosy is a chronic dermato-neurological disease caused by infection with Mycobacterium leprae. In 2013 almost 200,000 new cases of leprosy were detected around the world. Since the first symptoms take from years to decades to appear, the total number of asymptomatic patients is impossible to predict. Although leprosy is one of the oldest records of human disease, the mechanisms involved with its transmission and epidemiology are still not completely understood. In the present work, we experimentally investigated the hypothesis that the mosquitoes Aedes aegypti and Culex quinquefasciatus and the hemiptera Rhodnius prolixus act as leprosy vectors. By means of real-time PCR quantification of M. leprae 16SrRNA, we found that M. leprae remained viable inside the digestive tract of Rhodnius prolixus for 20 days after oral infection. In contrast, in the gut of both mosquito species tested, we were not able to detect M. leprae RNA after a similar period of time. Inside the kissing bug Rhodnius prolixus digestive tract, M. leprae was initially restricted to the anterior midgut, but gradually moved towards the hindgut, in a time course reminiscent of the life cycle of Trypanosoma cruzi, a well-known pathogen transmitted by this insect. The maintenance of M. leprae infectivity inside the digestive tract of this kissing bug is further supported by successful mice footpad inoculation with feces collected 20 days after infection. We conclude that Rhodnius prolixus defecate infective M. leprae, justifying the evaluation of the presence of M. leprae among sylvatic and domestic kissing bugs in countries endemic for leprosy. PMID:27203082
Neumann, Arthur da Silva; Dias, Felipe de Almeida; Ferreira, Jéssica da Silva; Fontes, Amanda Nogueira Brum; Rosa, Patricia Sammarco; Macedo, Rafael Enrique; Oliveira, José Henrique; Teixeira, Raquel Lima de Figueiredo; Pessolani, Maria Cristina Vidal; Moraes, Milton Ozório; Suffys, Philip Noel; Oliveira, Pedro L; Sorgine, Marcos Henrique Ferreira; Lara, Flavio Alves
Leprosy is a chronic dermato-neurological disease caused by infection with Mycobacterium leprae. In 2013 almost 200,000 new cases of leprosy were detected around the world. Since the first symptoms take from years to decades to appear, the total number of asymptomatic patients is impossible to predict. Although leprosy is one of the oldest records of human disease, the mechanisms involved with its transmission and epidemiology are still not completely understood. In the present work, we experimentally investigated the hypothesis that the mosquitoes Aedes aegypti and Culex quinquefasciatus and the hemiptera Rhodnius prolixus act as leprosy vectors. By means of real-time PCR quantification of M. leprae 16SrRNA, we found that M. leprae remained viable inside the digestive tract of Rhodnius prolixus for 20 days after oral infection. In contrast, in the gut of both mosquito species tested, we were not able to detect M. leprae RNA after a similar period of time. Inside the kissing bug Rhodnius prolixus digestive tract, M. leprae was initially restricted to the anterior midgut, but gradually moved towards the hindgut, in a time course reminiscent of the life cycle of Trypanosoma cruzi, a well-known pathogen transmitted by this insect. The maintenance of M. leprae infectivity inside the digestive tract of this kissing bug is further supported by successful mice footpad inoculation with feces collected 20 days after infection. We conclude that Rhodnius prolixus defecate infective M. leprae, justifying the evaluation of the presence of M. leprae among sylvatic and domestic kissing bugs in countries endemic for leprosy.
Bai, Chang-Ming; Wang, Qing-Chen; Morga, Benjamin; Shi, Jie; Wang, Chong-Ming
We investigated the susceptibility of ark shell, Scapharca broughtonii, adults to Ostreid herpesvirus SB strain (OsHV-1-SB) through experimental infection by intramuscular injection assays. Results showed the onset of mortality occurred at 3days post injection, one day after the water turbidity became evident in rearing tanks. The mortality curves for the challenged group were similar to those observed at affected hatcheries. Histological lesions, herpesvirus-like particles and high OsHV-1-SB quantities were detected in challenged ark shells. This is the first study to successfully reproduce OsHV-1 disease in Arcoida species, and very few studies in adult bivalves (over 24months old).
Nurelhuda, I E; Elowni, E E; Hassan, T
Oxfendazole was tested against Raillietina tetragona in experimentally infected chickens using single oral doses of 20, 10, 7.5, 5, and 2.5 mg/kg body weight. The minimal dose of the drug which produced 100% efficacy against immature worms was 10 mg/kg whereas the same effect on the mature parasite was obtained with 7.5 mg/kg. Doses lower than these significantly reduce worm burdens. The compound appears to be safe for chickens and a dose of 20 mg/kg (twice the recommended dose) produced no untoward clinical reactions.
Veljkovic, Veljko; Loiseau, Philippe M.; Figadere, Bruno; Glisic, Sanja; Veljkovic, Nevena; Perovic, Vladimir R.; Cavanaugh, David P.; Branch, Donald R.
The ongoing Ebola virus epidemic has presented numerous challenges with respect to control and treatment because there are no approved drugs or vaccines for the Ebola virus disease (EVD). Herein is proposed simple theoretical criterion for fast virtual screening of molecular libraries for candidate inhibitors of Ebola virus infection. We performed a repurposing screen of 6438 drugs from DrugBank using this criterion and selected 267 approved and 382 experimental drugs as candidates for treatment of EVD including 15 anti-malarial drugs and 32 antibiotics. An open source Web server allowing screening of molecular libraries for candidate drugs for treatment of EVD was also established. PMID:25717373
Vickers, M L; Hanson, R P
Red-winged blackbirds (Agelaius phoeniceus) were experimentally exposed to three avian paramyxoviruses: turkey parainfluenza virus, Yucaipa virus, and two strains of Newcastle disease virus. Aerosol exposure resulted in infection but exposure in food or drinking water rarely or never did. Tracheal swabs contained virus for up to eight days post exposure, cloacal swabs were negative. Transmission to contact birds occurred infrequently. Antibody response was of low titer and short duration. No hemagglutination inhibition activity against these viruses was found in 387 sera collected from red-winged blackbirds and tricolored blackbirds (Agelaius tricolor) trapped in six states.
Morphological and molecular investigations of a microsporidium infecting the European grape vine moth, Lobesia botrana Den. et Schiff., and its taxonomic determination as Cystosporogenes legeri nov. comb.
Kleespies, Regina G; Vossbrinck, Charles R; Lange, Martin; Jehle, Johannes A
We have isolated a microsporidium from a laboratory stock of the European grape vine moth, Lobesia botrana Den. et Schiff. (Lepidoptera, Tortricidae). Screening of this stock showed an infection rate of more than 90%, whereas field collected larvae from three different locations in Rhineland-Palatinate (Germany) did not demonstrate any signs of infection. Light and electron microscopic investigations of infected insects showed that gross pathology, morphology, and ultrastructure of the microsporidium are similar to those described earlier for Pleistophora legeri. Comparative phylogenetic analysis of the small subunit rDNA using maximum likelihood, maximum parsimony, and neighbour joining distance methods showed that our isolate was closely related to Cystosporogenes operophterae. Based on our morphological and molecular investigations we propose to rename this species Cystosporogenes legeri nov. comb.
Fourth European Conference on Infections in Leukaemia (ECIL-4): guidelines for diagnosis and treatment of human respiratory syncytial virus, parainfluenza virus, metapneumovirus, rhinovirus, and coronavirus.
Hirsch, Hans H; Martino, Rodrigo; Ward, Katherine N; Boeckh, Michael; Einsele, Hermann; Ljungman, Per
Community-acquired respiratory virus (CARV) infections have been recognized as a significant cause of morbidity and mortality in patients with leukemia and those undergoing hematopoietic stem cell transplantation (HSCT). Progression to lower respiratory tract infection with clinical and radiological signs of pneumonia and respiratory failure appears to depend on the intrinsic virulence of the specific CARV as well as factors specific to the patient, the underlying disease, and its treatment. To better define the current state of knowledge of CARVs in leukemia and HSCT patients, and to improve CARV diagnosis and management, a working group of the Fourth European Conference on Infections in Leukaemia (ECIL-4) 2011 reviewed the literature on CARVs, graded the available quality of evidence, and made recommendations according to the Infectious Diseases Society of America grading system. Owing to differences in screening, clinical presentation, and therapy for influenza and adenovirus, ECIL-4 recommendations are summarized for CARVs other than influenza and adenovirus.
Falcão de Oliveira, Everton; Oshiro, Elisa Teruya; Fernandes, Wagner de Souza; Murat, Paula Guerra; Medeiros, Márcio José de; Souza, Alda Izabel; Oliveira, Alessandra Gutierrez de; Galati, Eunice Aparecida Bianchi
Several parameters should be addressed before incriminating a vector for Leishmania transmission. Those may include its ability to become infected by the same Leishmania species found in humans, the degree of attractiveness for reservoirs and humans and capacity to sustain parasite infection under laboratory conditions. This study evaluated the vectorial capacity of Lutzomyia cruzi for Leishmania infantum and gathered information on its ability to harbor L. amazonensis. Laboratory-reared Lu. cruzi were infected experimentally by feeding them on dogs infected naturally with L. infantum and hamsters infected with L. amazonensis. Sand fly attractiveness to dogs and humans was determined using wild caught insects. The expected daily survival of infected Lu. cruzi, the duration of the gonotrophic cycle, and the extrinsic incubation period were also investigated for both parasites. Vector competence was investigated for both Leishmania species. The mean proportion of female sand flies that fed on hosts was 0.40. For L. infantum and L. amazonensis, Lu. cruzi had experimental infection rates of 10.55% and 41.56%, respectively. The extrinsic incubation period was 3 days for both Leishmania species, regardless of the host. Survival expectancy of females infected with L. infantum and L. amazonensis after completing the gonotrophic cycle was 1.32 and 0.43, respectively. There was no association between L. infantum infection and sand fly longevity, but L. amazonensis-infected flies had significantly greater survival probabilities. Furthermore, egg-laying was significantly detrimental to survival. Lu. cruzi was found to be highly attracted to both dogs and humans. After a bloodmeal on experimentally infected hosts, both parasites were able to survive and develop late-stage infections in Lu. cruzi. However, transmission was demonstrated only for L. amazonensis-infected sand flies. In conclusion, Lu. cruzi fulfilled several of the requirements of vectorial capacity for L. infantum
Murat, Paula Guerra; de Medeiros, Márcio José; Souza, Alda Izabel; de Oliveira, Alessandra Gutierrez
Several parameters should be addressed before incriminating a vector for Leishmania transmission. Those may include its ability to become infected by the same Leishmania species found in humans, the degree of attractiveness for reservoirs and humans and capacity to sustain parasite infection under laboratory conditions. This study evaluated the vectorial capacity of Lutzomyia cruzi for Leishmania infantum and gathered information on its ability to harbor L. amazonensis. Laboratory-reared Lu. cruzi were infected experimentally by feeding them on dogs infected naturally with L. infantum and hamsters infected with L. amazonensis. Sand fly attractiveness to dogs and humans was determined using wild caught insects. The expected daily survival of infected Lu. cruzi, the duration of the gonotrophic cycle, and the extrinsic incubation period were also investigated for both parasites. Vector competence was investigated for both Leishmania species. The mean proportion of female sand flies that fed on hosts was 0.40. For L. infantum and L. amazonensis, Lu. cruzi had experimental infection rates of 10.55% and 41.56%, respectively. The extrinsic incubation period was 3 days for both Leishmania species, regardless of the host. Survival expectancy of females infected with L. infantum and L. amazonensis after completing the gonotrophic cycle was 1.32 and 0.43, respectively. There was no association between L. infantum infection and sand fly longevity, but L. amazonensis–infected flies had significantly greater survival probabilities. Furthermore, egg-laying was significantly detrimental to survival. Lu. cruzi was found to be highly attracted to both dogs and humans. After a bloodmeal on experimentally infected hosts, both parasites were able to survive and develop late-stage infections in Lu. cruzi. However, transmission was demonstrated only for L. amazonensis–infected sand flies. In conclusion, Lu. cruzi fulfilled several of the requirements of vectorial capacity for L
De Brito, T; Gunji, J; Camargo, M E; Ceravolo, A; Da Silva, L C
Three monkeys (Cebus apella) experimentally infected with Schistosoma mansoni studied for periods of 19, 14, and 11 months showed deposits containing gamma-globulin in subendothelial and subepithelial basal membranes and in basement membranes proper. The glomeruli showed mild reactivity characterized by local hypertrophy and hyperplasia of mesangial cells. Such findings were close to those observed by us in the kidney of hepatosplenic schistosomiasis patients without evidence of renal disease. The distribution of the deposits, both in human and experimental disease, are suggestive of preformed, non-glomerular antigen-antibody complexes that form in a zone of excess antigen and become trapped in the glomerular capillaries.The possibility exists, but has not yet been proved beyond doubt, that renal disease in schistosomiasis patients could be the end result of this pathogenetic mechanism.
Suen, Willy W.; Uddin, Muhammad J.; Wang, Wenqi; Brown, Vienna; Adney, Danielle R.; Broad, Nicole; Prow, Natalie A.; Bowen, Richard A.; Hall, Roy A.; Bielefeldt-Ohmann, Helle
The economic impact of non-lethal human and equine West Nile virus (WNV) disease is substantial, since it is the most common presentation of the infection. Experimental infection with virulent WNV strains in the mouse and hamster models frequently results in severe neural infection and moderate to high mortality, both of which are not representative features of most human and equine infections. We have established a rabbit model for investigating pathogenesis and immune response of non-lethal WNV infection. Two species of rabbits, New Zealand White (Oryctolagus cuniculus) and North American cottontail (Sylvilagus sp.), were experimentally infected with virulent WNV and Murray Valley encephalitis virus strains. Infected rabbits exhibited a consistently resistant phenotype, with evidence of low viremia, minimal-absent neural infection, mild-moderate neuropathology, and the lack of mortality, even though productive virus replication occurred in the draining lymph node. The kinetics of anti-WNV neutralizing antibody response was comparable to that commonly seen in infected horses and humans. This may be explained by the early IFNα/β and/or γ response evident in the draining popliteal lymph node. Given this similarity to the human and equine disease, immunocompetent rabbits are, therefore, a valuable animal model for investigating various aspects of non-lethal WNV infections. PMID:26184326
Darwich, L; Li, Y; Serrano-Pérez, B; Mur-Novales, R; Garcia-Ispierto, I; Cabezón, O; López-Gatius, F; Almería, S
In the present study, IFN-γ (Th1), IL-17A (Th17) and IL-4 (Th2) concentrations in response to concanavalin (ConA) and Neospora caninum antigen (Nc-1) stimulation were determined in cultures of cells from control uninfected (n=4), naturally N. caninum-infected (n=3) and experimentally N. caninum-infected (n=6) pregnant dams and their foetuses. Experimental animals were infected at 110days of gestation and euthanized 6weeks post-infection. In culture supernatants from the dams, significantly higher IFN-γ and IL-4 levels were found in the experimentally-infected animals compared to the control or naturally-infected dams. However, among the experimentally-infected dams no significant differences in IFN-γ production were observed regardless of the incidence of live or aborted/dead foetuses, though spleen cultures of dams carrying live foetuses showed the highest levels of IFN-γ. IL-17A production was very low and occasional in the dams infected with N. caninum and did not seem to be a major regulator of IFN-γ production in this model. Experimentally infected dams with live foetuses showed higher IL-4 levels and accordingly IFN-γ/IL-4 ratios were significantly lower than ratios recorded for cows with aborted/dead foetuses. In the infected foetuses of these dams, only spleen cultures showed high levels of IFN-γ and IL-4 after Nc-1 antigen and ConA stimulation, respectively. No IL-17A was detected in the foetuses. As conclusion, although we could not clearly relate a protective immune response against N. caninum abortion only to IFN-γ levels in cell cultures, our results highlight the important role of an inverse IFN-γ/IL-4 balance in conferring protection against abortion induced by this parasite.
Geurden, T; Claerebout, E; Dursin, L; Deflandre, A; Bernay, F; Kaltsatos, V; Vercruysse, J
A controlled and blinded study was conducted to evaluate the efficacy and safety of a treatment with paromomycin sulphate against an experimental Giardia infection in calves. Animals were infected with 10(5)Giardia cysts of cattle origin and were either treated 11 days later with 25, 50 or 75 mg paromomycin/(kg body weight per day) during 5 consecutive days or not treated (control group). Efficacy was evaluated based on reduction in cyst excretion. Furthermore weight gain and diarrhea scores were monitored. In the group treated with 75 mg/kg per day there was a 100% reduction in cyst excretion until 9 days after the start of the treatment (D9) and a very high reduction (> or =98%) until D13. There was a high reduction (> or =93%) until D9 and D13 in the groups treated with 25 and 50 mg/kg, respectively. The cumulative cyst excretion on D13 was significantly (P<0.05) lower in the groups treated with 75 and 50 mg/kg compared to the control group. Although there was a trend towards higher weight gain and less diarrhea in the treated groups, differences between groups were not significant. No adverse reactions to the paromomycin treatment were recorded. Furthermore, the need for reliable parameters for evaluation of treatments against protozoal infections is emphasised.
Cruz-Rivera, Mayra; Vaughan, Gilberto; Mendlovic, Fela; Vergara-Castañeda, Arely; Romero-Valdovinos, Mirza; Leon-Cabrera, Sonia; Alonso, Monica; Avila, Guillermina; Flisser, Ana
The establishment of Taenia solium adult parasite in the human intestine causes taeniosis. Importantly, the immunological mechanisms occurring at the interface between the parasite and its host are not fully known. The development of experimental animal models has facilitated the understanding of the host-parasite relationship. In this study we standardized a quantitative RT-PCR method for analyzing hamster messenger RNA for interferon-gamma (IFN-γ) and interleukins (IL): IL-4 IL-10, IL-12 and IL-13. This method was then used to evaluate the local cytokine response elicited against the adult parasite at the attachment site in the intestine of infected hamsters. The results showed an intense IFN-γ response, as well as an up-regulation of IL-4 as early as three days post-infection, permanence of IL-10 until the end of the experiment and down regulation of IL-12. These data are in agreement with a bias toward a Th-2 response as the infection progresses.
Cannet, Arnaud; Akhoundi, Mohammad; Michel, Gregory; Marty, Pierre; Delaunay, Pascal
Leishmaniasis is a vector-borne disease that is transmitted by sandflies and caused by obligate intracellular protozoa of the genus Leishmania. In the present study, we carried out a screening on the experimental infection of Phlebotomus pernioucus by bioluminescent Leishmania infantum using murine model and artificial feeder. We developed a real-time polymerase chain reaction (RT-PCR)-based method to determine individually the number of Leishmania promastigotes fed by infected flies. Among 1840 new emerged female sand flies, 428 were fed on the infected mice. After their death, they were analysed individually by RT-PCR. Our results demonstrated just a single Leishmania positive female at sixth day post meal. A total of 1070 female sand flies were exposed in contact with artificial feeder containing the human blood with two different quantities of Leishmania parasites: 2.106/mL and 1.107/mL. A blood meal including 1.107/mL LUC-promastigotes was proposed to 270 females and 75 (28%) flies were engorged. Among them, 44 (59%) were positive by RT-PCR analysis, with a relative average of 50551 Leishmania parasites. In case of blood feeding of females with 2.106/mL promastigotes, 57 out of 800 (7%) females succeed to feed from artificial feeder which 22 (39%) were positive with a relative average of 6487 parasites. PMID:27439032
Rattanachaikunsopon, Pongsak; Phumkhachorn, Parichat
The essential oils extracted from the four herbs, cinnamon (Cinnamomum verum), clove (Syzygium aromaticum), ginger (Zingiber officinale) and holy basil (Ocimum sanctum), were investigated for their antimicrobial activity and mode of action against Lactococcus garvieae, a fish pathogenic bacteria causing lactococcosis. Of all the tested oils, clove oil had the strongest inhibitory effect and exhibited a bactericidal mode of action against the pathogenic bacterium. When an intraperitoneal infection of tilapia (Oreochromis niloticus) with L. garvieae was performed, the median lethal dose (LD(50)) was determined to be 1.78x10(2) CFU/fish. For an in vivo trial, no mortality was apparent in fish fed on the fish diets supplemented with 3% (w/w) of clove oil and with 0.5% (w/w) of oxytetracycline 5 d prior to the infection with L. garvieae. These results indicate that clove oil had a protective effect on experimental L. garvieae infection in tilapia and the potential to replace antibiotics for controlling the disease.
Metcalf, E S; Almond, G W; Routh, P A; Horton, J R; Dillman, R C; Orndorff, P E
The domestic pig, Sus scrofa domestica, was examined as a model for typhoid fever, a severe and systemic disease of humans caused by Salmonella typhi. Six pigs were inoculated 1 week post-weaning with approximately 10(10)colony forming units (cfu) of wild type Salmonella typhi strain ISP1820 intranasally and observed for 3 weeks. S. typhi was cultured from the tonsils of 50% of the pigs at necropsy. Cultures from all other organs analysed (ileum, colon, spleen and liver) were negative. No clinical or histopathological signs of disease were observed. Pigs inoculated in parallel with swine-virulent S. choleraesuis all exhibited signs of systemic salmonellosis indicating that the parameters of the experimental infection with S. typhi (e.g. route) were appropriate. Whereas the pig has a gastrointestinal tract that is very similar to humans, our results indicated that the unique features of host and microbe interaction needed to produce typhoid fever were not mimicked in swine. Nevertheless, our observation of tonsillar involvement was consistent with former observations of S. choleraesuis and S. typhimurium infections in swine and supports a role for the tonsil in all porcine salmonella infections.
Demiaszkiewicz, A W
Single doses (from 300 to 1000 larvi per an animal) of invasive larvae E. cervi Cameron, 1931, obtained from experimentally infected snails Helix pomatia L. were given to 17 guinea pigs and 17 golden hamsters. Clinical nervous symptoms in the form of paresis and paralysis of limbs occurred only in the guinea pigs which were given a dose of 1000 larvi. These animals died in the period from the 75th to 117th day of infection. From their central nervous system single adult males and females of E. cervi were isolated. In the lungs and mesenteries of 2 dead pigs live larvae of E. cervi were found. This fact proves that the guinea pig can fulfil the role of a final and a paratenic host of E. cervi. No clinical symptoms were noticed in any hamster. In hamsters dissected on the 7th day of infection live larvae of E. cervi were found in the mesentery and in the fleshy part of the diaphgram. After 14 days the larvae found both in the mesentery and in the diaphragm were dead and surrounded by cellular infiltration. A strong tissue reaction of the hamster after the administration of E. cervi larvae is responsible for the larvae destruction and resorption.
Chaturvedi, Ashok K; Hameed, Rumanasma S; Wozniak, Karen L; Hole, Camaron R; Leopold Wager, Chrissy M; Weintraub, Susan T; Lopez-Ribot, Jose L; Wormley, Floyd L
Cryptococcus gattii is a fungal pathogen that can cause life-threatening respiratory and disseminated infections in immune-competent and immune-suppressed individuals. Currently, there are no standardized vaccines against cryptococcosis in humans, underlying an urgent need for effective therapies and/or vaccines. In this study, we evaluated the efficacy of intranasal immunization with C. gattii cell wall associated (CW) and/or cytoplasmic (CP) protein preparations to induce protection against experimental pulmonary C. gattii infection in mice. BALB/c mice immunized with C. gattii CW and/or CP protein preparations exhibited a significant reduction in pulmonary fungal burden and prolonged survival following pulmonary challenge with C. gattii. Protection was associated with significantly increased pro-inflammatory and Th1-type cytokine recall responses, in vitro and increased C. gattii-specific antibody production in immunized mice challenged with C. gattii. A number of immunodominant proteins were identified following immunoblot analysis of C. gattii CW and CP protein preparations using sera from immunized mice. Immunization with a combined CW and CP protein preparation resulted in an early increase in pulmonary T cell infiltrates following challenge with C. gattii. Overall, our studies show that C. gattii CW and CP protein preparations contain antigens that may be included in a subunit vaccine to induce prolonged protection against pulmonary C. gattii infection.
Clark, Larry; Hall, Jeffrey; McLean, Robert; Dunbar, Michael; Klenk, Kaci; Bowen, Richard; Smeraski, Cynthia A
Populations of greater sage-grouse (Centrocercus urophasianus) have declined 45-80% in North America since 1950. Although much of this decline has been attributed to habitat loss, recent field studies have indicated that West Nile virus (WNV) has had a significant negative impact on local populations of grouse. We confirm the susceptibility of greater sage-grouse to WNV infection in laboratory experimental studies. Grouse were challenged by subcutaneous injection of WNV (10(3.2) plaque-forming units [PFUs]). All grouse died within 6 days of infection. The Kaplan-Meier estimate for 50% survival was 4.5 days. Mean peak viremia for nonvaccinated birds was 10(6.4) PFUs/ml (+/-10(0.2) PFUs/ml, standard error of the mean [SEM]). Virus was shed cloacally and orally. Four of the five vaccinated grouse died, but survival time was increased (50% survival=9.5 days), with 1 grouse surviving to the end-point of the experiment (14 days) with no signs of illness. Mean peak viremia for the vaccinated birds was 10(2.3) PFUs/ml (+/-10(0.6) PFUs/ml, SEM). Two birds cleared the virus from their blood before death or euthanasia. These data emphasize the high susceptibility of greater sage-grouse to infection with WNV.
da Silva Raposo, Ricardo; Santarém, Vamilton Alvares; Merigueti, Yslla Fernanda Fitz Balo; Rubinsky-Elefant, Guita; de Lima Cerazo, Letícia Maria; Pereira, Ludimilla; Zampieri, Bianca Pelegi; da Silva, Aristeu Vieira; Laposy, Cecília Braga
Toxocariasis is a geohelminth zoonosis with worldwide distribution, mainly transmitted through the ingestion of embryonated eggs of nematodes of the Toxocara genus. The disease can also be transmitted to humans as a result of eating raw or undercooked meat of paratenic hosts, such as chickens. Here, we standardized an enzyme-linked immunosorbent assay (ELISA) for evaluating experimentally the kinetic and avidity index (AI) of IgY in broiler chickens infected with different doses of Toxocara canis eggs (G1:100; G2: 1000; and G3: 5000; n = 12 per group). The test showed 91.7% sensitivity (CI 95%: 77.5-98.3) and 100% specificity (CI 95%: 92.6-100), and highest efficiency (97.0%) at 60 days post infection. Infection was characterized by the presence of high avidity antibodies in the chronic phase. Our results support that the ELISA can be a highly useful tool for the detection of anti-Toxocara antibodies in chickens.
Gall, A; Schirrmeier, H
Rabbit haemorrhagic disease (RHD) is usually a fatal disease in rabbits which has spread rapidly across the continents. While previous studies suggested persistence in rabbits to be an important factor in the epidemiology, the relevance of field virus infection of immune rabbits has not been investigated in experimentally infected animals before. This report describes for the first time the persistence of rabbit haemorrhagic disease virus (RHDV) genome for at least 15 weeks in rabbits immunized with an inactivated vaccine as well as a subunit vaccine and subsequently challenged with virulent RHDV. The viral RNA loads were determined by real-time reverse transcription-polymerase chain reaction. No conspicuous association of the detectable amount of RHDV RNA with the type of vaccine, the time after infection and--with one exception--the level of RHDV-specific antibodies in the immunized animals was observed. The results presented in this study are an urgent evidence for the existence of carrier animals as an important factor in the epidemiology of RHD.
Araújo, Adriano Fernando; de Oliveira, Gabriel; Vasconcelos, Juliana Fraga; Ersching, Jonatan; Dominguez, Mariana Ribeiro; Vasconcelos, José Ronnie; Machado, Alexandre Vieira; Gazzinelli, Ricardo Tostes; Bruna-Romero, Oscar; Soares, Milena Botelho; Rodrigues, Mauricio Martins
In earlier studies, we reported that a heterologous prime-boost regimen using recombinant plasmid DNA followed by replication-defective adenovirus vector, both containing Trypanosoma cruzi genes encoding trans-sialidase (TS) and amastigote surface protein (ASP) 2, provided protective immunity against experimental infection with a reticulotropic strain of this human protozoan parasite. Herein, we tested the outcome of genetic vaccination of F1 (CB10XBALB/c) mice challenged with myotropic parasite strains (Brazil and Colombian). Initially, we determined that the coadministration during priming of a DNA plasmid containing the murine IL-12 gene improved the immune response and was essential for protective immunity elicited by the heterologous prime-boost regimen in susceptible male mice against acute lethal infections with these parasites. The prophylactic or therapeutic vaccination of resistant female mice led to a drastic reduction in the number of inflammatory infiltrates in cardiac and skeletal muscles during the chronic phase of infection with either strain. Analysis of the electrocardiographic parameters showed that prophylactic vaccination reduced the frequencies of sinus arrhythmia and atrioventricular block. Our results confirmed that prophylactic vaccination using the TS and ASP-2 genes benefits the host against acute and chronic pathologies caused by T. cruzi and should be further evaluated for the development of a veterinary or human vaccine against Chagas disease. PMID:25061263
Gulyaeva, Marina A.; Zaykovskaia, Anna V.; Shestopalova, Lidia V.; Shestopalov, Aleksander M.
During 2006, H5N1 HPAI caused an epizootic in wild birds, resulting in a die-off of Laridae in the Novosibirsk region at Chany Lake. In the present study, we infected common gulls (Larus canus) with a high dose of the H5N1 HPAI virus isolated from a common gull to determine if severe disease could be induced over the 28 day experimental period. Moderate clinical signs including diarrhea, conjunctivitis, respiratory distress and neurological signs were observed in virus-inoculated birds, and 50% died. The most common microscopic lesions observed were necrosis of the pancreas, mild encephalitis, mild myocarditis, liver parenchymal hemorrhages, lymphocytic hepatitis, parabronchi lumen hemorrhages and interstitial pneumonia. High viral titers were shed from the oropharyngeal route and virus was still detected in one bird at 25 days after infection. In the cloaca, the virus was detected sporadically in lower titers. The virus was transmitted to direct contact gulls. Thus, infected gulls can pose a significant risk of H5N1 HPAIV transmission to other wild migratory waterfowl and pose a risk to more susceptible poultry species. These findings have important implications regarding the mode of transmission and potential risks of H5N1 HPAI spread by gulls. PMID:26243601
Chaturvedi, Ashok K.; Hameed, Rumanasma S.; Wozniak, Karen L.; Hole, Camaron R.; Leopold Wager, Chrissy M.; Weintraub, Susan T.; Lopez-Ribot, Jose L.; Wormley, Floyd L.
Cryptococcus gattii is a fungal pathogen that can cause life-threatening respiratory and disseminated infections in immune-competent and immune-suppressed individuals. Currently, there are no standardized vaccines against cryptococcosis in humans, underlying an urgent need for effective therapies and/or vaccines. In this study, we evaluated the efficacy of intranasal immunization with C. gattii cell wall associated (CW) and/or cytoplasmic (CP) protein preparations to induce protection against experimental pulmonary C. gattii infection in mice. BALB/c mice immunized with C. gattii CW and/or CP protein preparations exhibited a significant reduction in pulmonary fungal burden and prolonged survival following pulmonary challenge with C. gattii. Protection was associated with significantly increased pro-inflammatory and Th1-type cytokine recall responses, in vitro and increased C. gattii-specific antibody production in immunized mice challenged with C. gattii. A number of immunodominant proteins were identified following immunoblot analysis of C. gattii CW and CP protein preparations using sera from immunized mice. Immunization with a combined CW and CP protein preparation resulted in an early increase in pulmonary T cell infiltrates following challenge with C. gattii. Overall, our studies show that C. gattii CW and CP protein preparations contain antigens that may be included in a subunit vaccine to induce prolonged protection against pulmonary C. gattii infection. PMID:25119981
Lim, Stephanie M; Brault, Aaron C; van Amerongen, Geert; Bosco-Lauth, Angela M; Romo, Hannah; Sewbalaksing, Varsha D; Bowen, Richard A; Osterhaus, Albert D M E; Koraka, Penelope; Martina, Byron E E
West Nile virus (WNV) outbreaks in North America have been characterized by substantial die-offs of American crows (Corvus brachyrhynchos). In contrast, a low incidence of bird deaths has been observed during WNV epidemic activity in Europe. To examine the susceptibility of the western European counterpart of American crows, we inoculated carrion crows (Corvus corone) with WNV strains isolated in Greece (Gr-10), Italy (FIN and Ita09), and Hungary (578/10) and with the highly virulent North American genotype strain (NY99). We also inoculated American crows with a selection of these strains to examine the strains' virulence in a highly susceptible bird species. Infection with all strains, except WNV FIN, resulted in high rates of death and high-level viremia in both bird species and virus dissemination to several organs. These results suggest that carrion crows are highly susceptible to WNV and may potentially be useful as part of dead bird surveillance for early warning of WNV activity in Europe.
Lim, Stephanie M.; Brault, Aaron C.; van Amerongen, Geert; Bosco-Lauth, Angela M.; Romo, Hannah; Sewbalaksing, Varsha D.; Bowen, Richard A.; Osterhaus, Albert D.M.E.; Koraka, Penelope
West Nile virus (WNV) outbreaks in North America have been characterized by substantial die-offs of American crows (Corvus brachyrhynchos). In contrast, a low incidence of bird deaths has been observed during WNV epidemic activity in Europe. To examine the susceptibility of the western European counterpart of American crows, we inoculated carrion crows (Corvus corone) with WNV strains isolated in Greece (Gr-10), Italy (FIN and Ita09), and Hungary (578/10) and with the highly virulent North American genotype strain (NY99). We also inoculated American crows with a selection of these strains to examine the strains’ virulence in a highly susceptible bird species. Infection with all strains, except WNV FIN, resulted in high rates of death and high-level viremia in both bird species and virus dissemination to several organs. These results suggest that carrion crows are highly susceptible to WNV and may potentially be useful as part of dead bird surveillance for early warning of WNV activity in Europe. PMID:26197093
Sharma, Vikash; Jakhar, K. K.; Nehra, Vikas; Kumar, Sarvan
Aim: An experimental study was conducted on 192-day-old broiler chicks for evaluating the effect of 10% neem leaf extract (NLE) supplementationon biochemical parameters in chickens experimentally infected with Escherichia coli O78 at 107 CFU/0.5 ml at 7 days of age. Materials and Methods: The 192-day-old broiler chicks were procured. These chicks were divided into two groups (A and B) containing 96 birds each on the 1st day. Diet of all the chicks of Group A was supplemented with 10%NLE in water, whereas chicks of Group B were given feed and water devoid of NLE supplementation throughout the experiment. After rearing for 1 week, chicks of both the groups (A and B) were again divided into two subgroups (Group A into A1 and A2 and Group B into B1 and B2) of 54 and 42 birds, respectively. At the age of 7 days all the chicks of groups A1 and B1 were injected with E. coli O78 at 107 CFU/0.5 ml intraperitoneally. Blood samples were collected from six chicks from each group at day 0, 2, 4, 7, 14, 21, 28 days post-infection and serum was separated for biochemical studies. Results: There was a significant increase in serum alanine transaminase (ALT), aspartate transaminase (AST), lactate dehydrogenase (LDH) activities, globulin concentration and a decrease in total protein (TP), albumin concentrations, and alkaline phosphatase (ALP) activity in both the infected groups. However, the changes in biochemical values, i.e., ALT, AST, LDH, ALP, TP, albumin, and globulin wereof lower magnitude in NLE supplemented group suggesting hepatoprotective and cardioprotective effect of NLE. Conclusions: Fromthe present study, it is reasonable to conclude that significant increase in the value of ALT, AST, LDH, globulin, and significant decrease in the value of ALP, TP, and albumin was of lower magnitude in supplemented infected group (A1) as compared to non-supplemented infected group (B1) suggesting hepatoprotective and cardioprotective effect of NLE. PMID:27047040
Segarra, Amélie; Baillon, Laury; Faury, Nicole; Tourbiez, Delphine; Renault, Tristan
High mortality rates are reported in spat and larvae of Pacific oyster Crassostrea gigas and associated with ostreid herpesvirus 1 (OsHV-1) detection in France. Although the viral infection has been experimentally reproduced in oyster larvae and spat, little knowledge is currently available concerning the viral entry and its distribution in organs and tissues. This study compares OsHV-1 DNA and RNA detection and localization in experimentally infected oysters using two virus doses: a low dose that did not induce any mortality and a high dose inducing high mortality. Real time PCR demonstrated significant differences in terms of viral DNA amounts between the two virus doses. RNA transcripts were detected in oysters receiving the highest dose of viral suspension whereas no transcript was observed in oysters injected with the low dose. This study also allowed observing kinetics of viral DNA and RNA detection in different tissues of oyster spat. Finally, viral detection was significantly different in function of tissues (p<0.005), time (p<0.005) with an interaction between tissues and time (p<0.005) for each probe.
Ito, A; Sakakibara, Y; Ma, L; Asano, K; Takiguchi, M; Yasuda, J; Hashimoto, A
Rats experimentally infected with Taenia taeniaeformis were followed-up until 14 weeks post inoculation with eggs (PIE) by hepatic ultrasonographic (US) image and serum antibody response analyses. Parasitic cysts could be imaged as small (2 mm in diameter) anechoic areas with or without a parenthesis-like echogenic small line from two weeks PIE. Immunoblot analysis using antigens from oncospheres (TtO), 30-day-old (TtM-30) and 300-day-old metacestodes (TtM-300) revealed that: (1) these three different developmental stages showed their own unique patterns suggesting the presence of stage-specific antigens; (2) faint IgM antibody responses to some components of TtO and TtM-30 or TtM-300 could be detected from one and two weeks PIE, respectively, and (3) IgG responses to some major components of both TtO and TtM-300, and TtM-30 were easily detected from four and five weeks PIE onwards, respectively. Both TtO and TtM (especially TtM-300) appeared to be highly useful for detection of antibody responses in experimentally infected rats. Due to the easiness in preparation of antigens, fully developed metacestodes may be the best candidate antigens for serodiagnosis. These results strongly suggest that both US image and antibody analyses using antigens from fully developed metacestodes are useful for detection of the early stage of cysticercosis in laboratory animal model.
King, T A; Cable, J
Parasites represent a threat to endangered fish species, particularly when the parasite can host switch and the new host is vulnerable. If the parasite is highly host specific then successful host switching should be a rare occurrence; however, the host range of many parasites which are assumed to be specialists has never been tested. This includes the monogenean Gyrodactylus turnbulli, a well-studied ectoparasite found caudally on its known host, the guppy, Poecilia reticulata. In this study, we monitored parasite establishment and reproduction on a range of poeciliids and more distantly related fish. Individually maintained fish were experimentally infected with a single parasite and monitored daily to establish whether G. turnbulli could survive and reproduce on other fish species. Gyrodactylus turnbulli can infect a wider range of hosts than previously considered, highlighting the fact that host specificity can never be assumed unless experimentally tested. Our findings also have significant implications for parasite transmission to novel hosts and provide further insight into the evolutionary origins of this ubiquitous group of fish pathogens. Previous molecular evidence indicates that host switching is the key mechanism for speciation within the genus Gyrodactylus. Until recently, most Gyrodactylus spp. were assumed to be narrowly host specific. However, our findings suggest that even so-called specialist species, such as G. turnbulli, may represent a threat to vulnerable fish stocks. In view of the potential importance of host switching under artificial conditions, we propose to describe this as 'artificial ecological transfer' as opposed to 'natural ecological transfer', host switching under natural conditions.
Tell, Lisa A; Clemons, Karl V; Kline, Yvonne; Woods, Leslie; Kass, Philip H; Martinez, Marife; Stevens, David A
Aspergillus fumigatus causes disease in birds. Our objectives were to determine pharmacokinetic parameters and evaluate efficacy of voriconazole (VCZ) in a novel experimental quail model. After a single oral VCZ dose of 20 or 40 mg/kg, plasma concentrations peaked 2 h postdose (5.8 and 6.9 microg/ml) and remained above 0.5 microg/ml for 4 and 12 h postdose, respectively. For the efficacy study, ten-day-old Japanese quail (n = 60) were infected intratracheally with A. fumigatus conidia. Daily oral VCZ at 20 or 40 mg/kg was initiated 24 h postinfection (PI); infected diluent-treated birds were given de-ionized water orally. Preassigned birds were euthanized on days 5 or 10 PI. VCZ at 40 mg/kg prolonged survival compared to 20 mg/kg or diluent-treatment (P < 0.05) and lungs from birds given VCZ at 40 mg/kg had fewer colony forming units (CFU) than diluent-treated (P = 0.03). At day 10 PI, birds treated with VCZ at 20 mg/kg had significantly fewer fungi in the lungs as demonstrated by methenamine silver stain (P = 0.017) or immunohistochemistry, as compared to diluent-treated (P = 0.034). Histopathologically, VCZ-treated birds did not have necrotic lesions and showed a trend toward fewer with acute inflammatory changes. VCZ at 40 mg/kg was efficacious in quail with experimental pulmonary aspergillosis.
Hall, Jeffrey S.; Krauss, Scott; Franson, J. Christian; TeSlaa, Joshua L.; Nashold, Sean W.; Stallknecht, David E.; Webby, Richard J.; Webster, Robert G.
Background: Low pathogenic avian influenza viruses (LPAIV) have been reported in shorebirds, especially at Delaware Bay, USA, during spring migration. However, data on patterns of virus excretion, minimal infectious doses, and clinical outcome are lacking. The ruddy turnstone (Arenaria interpres) is the shorebird species with the highest prevalence of influenza virus at Delaware Bay. Objectives: The primary objective of this study was to experimentally assess the patterns of influenza virus excretion, minimal infectious doses, and clinical outcome in ruddy turnstones. Methods: We experimentally challenged ruddy turnstones using a common LPAIV shorebird isolate, an LPAIV waterfowl isolate, or a highly pathogenic H5N1 avian influenza virus. Cloacal and oral swabs and sera were analyzed from each bird. Results: Most ruddy turnstones had pre-existing antibodies to avian influenza virus, and many were infected at the time of capture. The infectious doses for each challenge virus were similar (103·6–104·16 EID50), regardless of exposure history. All infected birds excreted similar amounts of virus and showed no clinical signs of disease or mortality. Influenza A-specific antibodies remained detectable for at least 2 months after inoculation. Conclusions: These results provide a reference for interpretation of surveillance data, modeling, and predicting the risks of avian influenza transmission and movement in these important hosts.
Truong, Thang; Boshra, Hani; Embury-Hyatt, Carissa; Nfon, Charles; Gerdts, Volker; Tikoo, Suresh; Babiuk, Lorne A.; Kara, Pravesh; Chetty, Thireshni; Mather, Arshad; Wallace, David B.; Babiuk, Shawn
Peste des petits ruminants (PPR) is a viral disease which primarily affects small ruminants, causing significant economic losses for the livestock industry in developing countries. It is endemic in Saharan and sub-Saharan Africa, the Middle East and the Indian sub-continent. The primary hosts for peste des petits ruminants virus (PPRV) are goats and sheep; however recent models studying the pathology, disease progression and viremia of PPRV have focused primarily on goat models. This study evaluates the tissue tropism and pathogenesis of PPR following experimental infection of sheep and goats using a quantitative time-course study. Upon infection with a virulent strain of PPRV, both sheep and goats developed clinical signs and lesions typical of PPR, although sheep displayed milder clinical disease compared to goats. Tissue tropism of PPRV was evaluated by real-time RT-PCR and immunohistochemistry. Lymph nodes, lymphoid tissue and digestive tract organs were the predominant sites of virus replication. The results presented in this study provide models for the comparative evaluation of PPRV pathogenesis and tissue tropism in both sheep and goats. These models are suitable for the establishment of experimental parameters necessary for the evaluation of vaccines, as well as further studies into PPRV-host interactions. PMID:24498032
Busatti, Haendel G N O; Alves, Ricardo J; Santana-Anjos, Karla G; Gil, Frederico F; Cury, Marcia C; Vannier-Santos, Marcos A; Gomes, Maria A
The chemotherapeutic agents used for the treatment of giardiasis are often associated with adverse side effects and are refractory cases, due to the development of resistant parasites. Therefore the search for new drugs is required. We have previously reported the giardicidal effects of metronidazole (MTZ) and its analogues (MTZ-Ms, MTZ-Br, MTZ-N(3), and MTZ-I) on the trophozoites of Giardia lamblia. Now we evaluated the activity of some giardicidal MTZ analogues in experimental infections in gerbils and its effects on the morphology and ultrastructural organization of Giardia. The giardicidal activity in experimental infections showed ED(50) values significantly lower for MTZ-I and MTZ-Br when compared to MTZ. Transmission electron microscopy was employed to approach the mechanism(s) of action of MTZ analogues upon the protozoan. MTZ analogues were more active than MTZ in changing significantly the morphology and ultrastructure of the parasite. The analogues affected parasite cell vesicle trafficking, autophagy, and triggered differentiation into cysts. These results coupled with the excellent giardicidal activity and lower toxicity demonstrate that these nitroimidazole derivates may be important therapeutic alternatives for combating giardiasis. In addition, our results suggest a therapeutic advantage in obtaining synthetic metronidazole analogues for screening of activities against other infectious agents.
Truong, Thang; Boshra, Hani; Embury-Hyatt, Carissa; Nfon, Charles; Gerdts, Volker; Tikoo, Suresh; Babiuk, Lorne A; Kara, Pravesh; Chetty, Thireshni; Mather, Arshad; Wallace, David B; Babiuk, Shawn
Peste des petits ruminants (PPR) is a viral disease which primarily affects small ruminants, causing significant economic losses for the livestock industry in developing countries. It is endemic in Saharan and sub-Saharan Africa, the Middle East and the Indian sub-continent. The primary hosts for peste des petits ruminants virus (PPRV) are goats and sheep; however recent models studying the pathology, disease progression and viremia of PPRV have focused primarily on goat models. This study evaluates the tissue tropism and pathogenesis of PPR following experimental infection of sheep and goats using a quantitative time-course study. Upon infection with a virulent strain of PPRV, both sheep and goats developed clinical signs and lesions typical of PPR, although sheep displayed milder clinical disease compared to goats. Tissue tropism of PPRV was evaluated by real-time RT-PCR and immunohistochemistry. Lymph nodes, lymphoid tissue and digestive tract organs were the predominant sites of virus replication. The results presented in this study provide models for the comparative evaluation of PPRV pathogenesis and tissue tropism in both sheep and goats. These models are suitable for the establishment of experimental parameters necessary for the evaluation of vaccines, as well as further studies into PPRV-host interactions.
Hausmann, Jennifer C; Wack, Allison N; Allender, Matthew C; Cranfield, Mike R; Murphy, Kevin J; Barrett, Kevin; Romero, Jennell L; Wellehan, James F X; Blum, Stella A; Zink, M Christine; Bronson, Ellen
The Maryland Zoo in Baltimore experienced an outbreak of Frog virus-3 (FV3)-like ranavirus during the summer of 2011, during which 14 of 27 (52%) of its captive eastern box turtles (Terrapene carolina carolina) survived. To assess survival, immunity, and viral shedding, an experimental challenge study was performed in which the surviving, previously infected turtles were reinfected with the outbreak strain of FV3-like ranavirus. Seven turtles were inoculated with virus intramuscularly and four control turtles received saline intramuscularly. The turtles were monitored for 8 wk with blood and oral swabs collected for quantitative polymerase chain reaction (qPCR). During that time, one of seven (14%) inoculated turtles and none of the controls (0%) died; there was no significant difference in survival. Clinical signs of the inoculated turtles, except for the turtle that died, were mild compared to the original outbreak. Quantitative PCR for FV3-like ranavirus on blood and oral swabs was positive for all inoculated turtles and negative for all controls. The turtle that died had intracytoplasmic inclusion bodies in multiple organs. Three inoculated and two control turtles were euthanized at the end of the study. No inclusion bodies were present in any of the organs. Quantitative PCR detected FV3-like ranavirus in the spleen of a control turtle, which suggested persistence of the virus. The surviving five turtles were qPCR-negative for FV3-like ranavirus from blood and oral swabs after brumation. Quantitative PCR for Terrapene herpesvirus 1 found no association between ranavirus infection and herpesvirus loads. In conclusion, previously infected eastern box turtles can be reinfected with the same strain of FV3-like ranavirus and show mild to no clinical signs but can shed the virus from the oral cavity.
Gast, Richard K; Guraya, Rupa; Guard, Jean
The continuing attribution of human Salmonella Enteritidis infections to internally contaminated eggs has necessitated the commitment of substantial public and private resources to Salmonella Enteritidis testing and control programs in commercial laying flocks. Cost-effective risk-reduction requires a detailed and comprehensive understanding of how Salmonella Enteritidis infections in hens result in deposition of the pathogen inside eggs. The present study sought to resolve some incompletely defined aspects of the relationship between Salmonella Enteritidis oral-exposure dose levels in experimentally infected laying hens and the frequency and location of subsequent egg contamination. In two trials, groups of specific-pathogen-free hens were experimentally inoculated with oral doses of 10(4), 10(6), or 10(8) CFU of a phage type 4 Salmonella Enteritidis strain. Eggs were collected 5 to 23 days postinoculation, and the yolk and albumen of each egg were cultured separately to detect Salmonella Enteritidis contamination. Larger oral doses of Salmonella Enteritidis administered to hens were associated with significant increases in the frequencies of both yolk and albumen contamination. Moreover, Salmonella Enteritidis was found in the albumen of a far-higher proportion of contaminated eggs from hens given the largest dose than from the other two groups. Salmonella Enteritidis contamination was detected in 0.7% of yolk and 0.2% of albumen samples after inoculation of hens with 10(4) CFU, 4.0% of yolk and 1.7% of albumen samples after inoculation with 10(6) CFU, and 6.5% of yolk and 10.8% of albumen samples after inoculation with 10(8) CFU. These results demonstrate that oral-exposure doses of Salmonella Enteritidis for laying hens can significantly affect both the frequency and location of deposition of this pathogen inside eggs.
Isaäcson, Margaretha; Taylor, Paul; Arntzen, Lorraine
The Mastomys natalensis species complex, subdivided into genetically distinct species having diploid chromosome numbers 2n = 32 and 2n = 36, is a reservoir for several zoonoses including Lassa fever and plague. This report describes a study to determine whether these sibling species and three other rodent species have different potential as reservoirs for plague. It was found that M. natalensis (2n = 32) was significantly more resistant to experimental plague infection (50% survived inoculation with 120 000 Yersinia pseudotuberculosis subsp. pestis) than was M. coucha (2n = 36) (none of which survived doses of 190 Y. pseudotuberculosis subsp.pestis). In descending order of resistance were M. natalensis, Aethomys chrysophilus, M. coucha, Tatera leucogaster and A. namaquensis. No A. namaquensis survived inoculation of 10 or more plague bacilli. Previous reports on susceptibility to plague or other infections, which were based exclusively on findings in the universally distributed laboratory-bred Mastomys, are thus not necessarily applicable to the M. natalensis species as a whole but probably only to M. coucha. The Y. pseudotuberculosis subsp. pestis fraction-1 passive haemagglutination test appeared to be relatively insensitive in that only 5 out of 47 animals surviving experimental plague infection showed specific antibodies 6 weeks after challenge. The geographic distribution of human plague in southern Africa corresponds closely with that of the plague-susceptible species, M. coucha, while the resistant species, M. natalensis, predominates in areas where human plague has not been recorded. The role of A. namaquensis in the ecology of plague needs to be carefully studied and its possible importance in plague research should be investigated further. PMID:6345015
Isaäcson, M; Taylor, P; Arntzen, L
The Mastomys natalensis species complex, subdivided into genetically distinct species having diploid chromosome numbers 2n = 32 and 2n = 36, is a reservoir for several zoonoses including Lassa fever and plague. This report describes a study to determine whether these sibling species and three other rodent species have different potential as reservoirs for plague. It was found that M. natalensis (2n = 32) was significantly more resistant to experimental plague infection (50% survived inoculation with 120 000 Yersinia pseudotuberculosis subsp. pestis) than was M. coucha (2n = 36) (none of which survived doses of 190 Y. pseudotuberculosis subsp.pestis). In descending order of resistance were M. natalensis, Aethomys chrysophilus, M. coucha, Tatera leucogaster and A. namaquensis. No A. namaquensis survived inoculation of 10 or more plague bacilli.Previous reports on susceptibility to plague or other infections, which were based exclusively on findings in the universally distributed laboratory-bred Mastomys, are thus not necessarily applicable to the M. natalensis species as a whole but probably only to M. coucha. The Y. pseudotuberculosis subsp. pestis fraction-1 passive haemagglutination test appeared to be relatively insensitive in that only 5 out of 47 animals surviving experimental plague infection showed specific antibodies 6 weeks after challenge.The geographic distribution of human plague in southern Africa corresponds closely with that of the plague-susceptible species, M. coucha, while the resistant species, M. natalensis, predominates in areas where human plague has not been recorded. The role of A. namaquensis in the ecology of plague needs to be carefully studied and its possible importance in plague research should be investigated further.
Ozwara, Hastings; Langermans, Jan A M; Maamun, Jenneby; Farah, Idle O; Yole, Dorcas S; Mwenda, Jason M; Weiler, Horst; Thomas, Alan W
Experimental systems that model some of the complex interactions between parasite and host can be extremely valuable in identifying and developing new prophylactics and therapeutics against human diseases. Because primates have similar immune systems to humans, we have characterized a baboon model for understanding host response to Plasmodium knowlesi. Ten intact olive baboons (Papio anubis) of either sex were experimentally infected with P. knowlesi H strain erythrocytic parasites. The infection in these baboons was either acute or chronic. Animals with acute infection developed multiple system organ dysfunction and cerebral involvement. In chronically infected animals, only the spleen was moderately enlarged. The P. knowlesi parasitemia profile in baboons and rhesus monkeys was comparable. However, some clinical symptoms of the baboons and P. falciparum-infected humans were similar. These studies demonstrate for the first time that P. anubis is a suitable host for P. knowlesi for studying clinical symptoms and pathology.
Pope, Robert A.; Parida, Satya; Bailey, Dalan; Brownlie, Joe; Barrett, Thomas; Banyard, Ashley C.
Peste-des-petits ruminants virus (PPRV) is a viral pathogen that causes a devastating plague of small ruminants. PPRV is an economically significant disease that continues to be a major obstacle to the development of sustainable agriculture across the developing world. The current understanding of PPRV pathogenesis has been heavily assumed from the closely related rinderpest virus (RPV) and other morbillivirus infections alongside data derived from field outbreaks. There have been few studies reported that have focused on the pathogenesis of PPRV and very little is known about the processes underlying the early stages of infection. In the present study, 15 goats were challenged by the intranasal route with a virulent PPRV isolate, Côte d’Ivoire ’89 (CI/89) and sacrificed at strategically defined time-points post infection to enable pre- and post-mortem sampling. This approach enabled precise monitoring of the progress and distribution of virus throughout the infection from the time of challenge, through peak viraemia and into a period of convalescence. Observations were then related to findings of previous field studies and experimental models of PPRV to develop a clinical scoring system for PPRV. Importantly, histopathological investigations demonstrated that the initial site for virus replication is not within the epithelial cells of the respiratory mucosa, as has been previously reported, but is within the tonsillar tissue and lymph nodes draining the site of inoculation. We propose that virus is taken up by immune cells within the respiratory mucosa which then transport virus to lymphoid tissues where primary virus replication occurs, and from where virus enters circulation. Based on these findings we propose a novel clinical scoring methodology for PPRV pathogenesis and suggest a fundamental shift away from the conventional model of PPRV pathogenesis. PMID:23418464
Dietz, Stefanie; Mack, Sarah-Lena; Hoelzle, Katharina; Becker, Katja; Jannasch, Carolin; Stadler, Julia; Ritzmann, Mathias; Hoelzle, Ludwig E
The uncultivable hemotrophic bacterium Mycoplasma suis causes infectious anemia in pigs worldwide. The mechanisms by which M. suis is transmitted from pig to pig are largely unknown. Thus, the present study aimed at investigating urine, feces, saliva, nasal and vaginal secrets as well as environmental samples for the presence of M. suis DNA to get insights into potential transmission routes. Seven pigs were experimentally infected with M. suis KI3806. Samples were taken for 8 days post infection (p.i.). A quantitative LightCycler msg1 PCR was used to detect and quantify M. suis. Shedding was found in saliva as well as nasal and vaginal secrets from day 6 p.i. on with a quantity of 3.4 × 10(2) to 2.7 × 10(5)M. suis/swab. In urine M. suis DNA could be detected in 100.0% of the samples from day 6 p.i. on with a quantity of 4.7 × 10(2) to 6.3 × 10(5)M. suis per mL. When shedding patterns were correlated to the median bacterial blood loads shedding was observed at loads of 2.0 × 10(9)-7.0 × 10(10)M. suis per mL blood. No M. suis DNA could be amplified from feces. Dust and water samples of the pig drinking troughs were positive for M. suis on days 2 and 6 post infection, air samples were M. suis-negative throughout the experiment. Our results indicate that blood independent direct transmission as well as indirect transmission via environmental contamination could play a role in the epidemiology of M. suis infections.
de Azevedo, Maria Isabel; Ferreiro, Laerte; Da Silva, Aleksandro S; Tonin, Alexandre A; Thorstenberg, Maria Luiza; Catilhos, Livia Gelain; França, Raqueli T; Leal, Daniela B R; Duarte, Marta M M F; Lopes, Sonia T A; Sangoi, Manuela B; Moresco, Rafael N; Fighera, Rafael; Santurio, Janio M
The aim of this study was to assess the role of the acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) as biomarkers of inflammation and tissue injury on rats experimentally infected by Cryptococcus neoformans. For this purpose, 20 male rats were divided into two groups: 10 animals representing the uninfected control group (Group A) and 10 C. neoformans var. grubii infected animals (Group B). Blood and brain samples were collected on days 10 (A10 and B10), and 30 (A30 and B30) post-infection (PI) for hematological analyses; AChE (in lymphocytes and brain) and seric BChE activity; interleukins (IL-1, IL-6, and IL-10); nitrite/nitrate (NOx) levels; and markers of protein oxidation (AOPP) and lipid peroxidation (TBARS). As a result, when animals of Group A were compared to animals of Group B, it was observed leukocytosis (P<0.05) on day 10 PI; AChE activity increase (P<0.05) in lymphocytes (day 30 PI) and in brain (days 10 and 30 PI); BChE activity decrease (P<0.05) on day 10 PI; IL-1 and IL-6 increase (P<0.01) in both periods, while IL-10 had reduced levels (P<0.01) in the same periods; NOx levels increased (P<0.05) significantly on days 10 and 30 PI, while AOPP and TBARS levels increased significantly on day 30 PI; as well as pneumonia on infected rats. Therefore, based on the results obtained, it was possible to conclude that AChE and BChE behavior lead to a proinflammatory reaction evidenced by the enhancement of IL-1, IL-6, and NOx throughout the experiment associated with reduction on IL-10 levels, and cellular damage.
El-Sabaa, Abdel-Aleem A.
Trichinosis is a parasitic zoonosis caused by the nematode Trichinella spiralis. Anthelmintics are used to eliminate intestinal adults as well as tissue-migrating and encysted larvae. This study aimed to investigate the effects of ivermectin and myrrh obtained from the aloe-gum resin of Commiphora molmol on experimental trichinosis. Ninety albino mice were orally infected with 300 T. spiralis larvae. Drugs were tested against adult worms at day 0 and day 5 and against encysted larvae on day 15 and day 35 post-infection (PI). Mature worms and encysted larvae were counted in addition to histopathological examination of muscle specimens. Serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), total protein, albumin, globulin, urea, and creatinine values were estimated. Significant reductions in mean worm numbers were detected in ivermectin treated mice at day 0 and day 5 PI achieving efficacies of 98.5% and 80.0%, while efficacies of myrrh in treated mice were 80.7% and 51.5%, respectively. At days 15 and 35 post-infection, ivermectin induced significant reduction in encysted larval counts achieving efficacies of 76.5% and 54.0%, respectively, while myrrh efficacies were 76.6% and 35.0%, respectively. AST, ALT, urea, and creatinine levels were reduced, while total proteins were increased in response to both treatments compared to their values in the infected non-treated mice. Ivermectin use for controlling T. spiralis could be continued. Myrrh was effective and could be a promising drug against the Egyptian strains of T. spiralis with results nearly comparable to ivermectin. PMID:23864740
Stuen, Snorre; Scharf, Wiebke; Schauer, Sonja; Freyburger, Felix; Bergström, Karin; von Loewenich, Friederike D
Anaplasma phagocytophilum transmitted by Ixodes spp. ticks is the causative agent of tick-borne fever (TBF) in domestic ruminants. TBF is widespread along the coast of southern Norway and may cause a severe problem for the sheep industry. Red deer (Cervus elaphus) are important hosts for ticks and have been found to be infected naturally with A. phagocytophilum. However, it is unclear whether red deer could serve as reservoir hosts for A. phagocytophilum infections in sheep. We infected lambs experimentally with a red deer and a sheep isolate, respectively. The 497 base pairs of the partial 16S rRNA gene sequences of both isolates were 100% identical to GenBank accession number M73220; the 3.8 kilobases of the total ank gene sequences were 99% identical. Sixteen lambs were used, four lambs in each group. Two groups were inoculated with the red deer isolate on day 0, and then challenged on day 42 with the ovine or the red deer isolate, respectively. The third group was inoculated with the sheep isolate on day 0 and challenged with the red deer strain on day 42. Four lambs were used as uninfected controls. Blood samples for hematology, bacteriology, and serology were collected regularly for 12 wk. Presence of A. phagocytophilum in blood was determined using blood smears. Serologic response was measured by indirect immunofluorescence. Although animals inoculated with the ovine strain showed more severe clinical manifestations, lambs infected with the red deer isolate reacted with typical signs of TBF such as fever, bacteremia, and neutropenia. We conclude that A. phagocytophilum strains causing TBF in sheep might circulate in the red deer population in Norway.
Ding, H; Wang, L; Shen, X; Gu, X; Zeng, D; Zeng, Z
The plasma and tissue pharmacokinetics of marbofloxacin in chickens experimentally infected with Mycoplasma gallisepticum and Escherichia coli were studied. Marbofloxacin was given to 66 infected chickens by oral administration at a dosage of 5 mg/kg b.w., once a day for three days. Plasma, brain, kidney, liver, lung, muscle and trachea were collected and marbofloxacin concentrations were analyzed by a high performance liquid chromatography method. In the infected chickens, maximal marbofloxacin concentrations in plasma, brain, kidney, liver, lung, muscle and trachea were 1.84, 1.33, 7.35, 5.61, 3.12, 2.98, and 4.51 g/mL (g); the elimination half-lives of marbofloxacin were 6.8, 2.74, 9.31, 8.45, 9.55, 11.53 and 5.46 h for plasma, brain, kidney, liver, lung, muscle and trachea, respectively. AUC were calculated to be 9.68, 8.04, 45.1, 27.03, 20.56, 19.47, and 32.68 μg/mL (g) for plasma, brain, kidney, liver, lung, muscle and trachea, respectively. Marbofloxacin concentration in tissues except for brain exceeded marbofloxacin concentration in plasma, with AUC(tissue) /AUC(plasma) ranging from 2.01 to 4.66 and Peak(tissue) /Peak(plasma) ranging from 1.62 to 3.99. The results showed that a marbofloxacin dosage of 5 mg/kg administered orally at 24 h intervals may provide successful treatment of chicken with MG and E. coli infection.
Yoneda, Misako; Guillaume, Vanessa; Sato, Hiroki; Fujita, Kentaro; Georges-Courbot, Marie-Claude; Ikeda, Fusako; Omi, Mio; Muto-Terao, Yuri; Wild, T Fabian; Kai, Chieko
Nipah virus (NiV) P gene encodes P protein and three accessory proteins (V, C and W). It has been reported that all four P gene products have IFN antagonist activity when the proteins were transiently expressed. However, the role of those accessory proteins in natural infection with NiV remains unknown. We generated recombinant NiVs lacking V, C or W protein, rNiV(V-), rNiV(C-), and rNiV(W-), respectively, to analyze the functions of these proteins in infected cells and the implications in in vivo pathogenicity. All the recombinants grew well in cell culture, although the maximum titers of rNiV(V-) and rNiV(C-) were lower than the other recombinants. The rNiV(V-), rNiV(C-) and rNiV(W-) suppressed the IFN response as well as the parental rNiV, thereby indicating that the lack of each accessory protein does not significantly affect the inhibition of IFN signaling in infected cells. In experimentally infected golden hamsters, rNiV(V-) and rNiV(C-) but not the rNiV(W-) virus showed a significant reduction in virulence. These results suggest that V and C proteins play key roles in NiV pathogenicity, and the roles are independent of their IFN-antagonist activity. This is the first report that identifies the molecular determinants of NiV in pathogenicity in vivo.
de Azevedo, Maria Isabel; Ferreiro, Laerte; Da Silva, Aleksandro S; Tonin, Alexandre A; Ruchel, Jader B; Rezer, João F P; França, Raqueli T; Zimmermann, Carine E P; Leal, Daniela B R; Duarte, Marta M M F; Lopes, Sonia T A; Flores, Mariana M; Fighera, Rafael; Santurio, Janio M
Cryptococcus neoformans, the etiological agent of cryptococcosis, is an opportunistic fungal pathogen of immunocompromised individuals. The aim of this study was to evaluate the activities of E-NTPDase and E-ADA in rats experimentally infected by C. neoformans var. grubii. Adult rats (35) were divided in two groups: 18 for the control group (uninfected) (A), and 17 for the infected group (B). Each group was separated into three sub-groups (A1, A2, A3-B1, B2, B3), and samples were collected on 10, 20, and 30 days post-infection (PI). Leukocyte counts, IFN-γ, TNF-α, IgM, IgG levels, and E-NTPDase and E-ADA activities were analyzed. It was possible to observe that IgG and IgM seric levels of infected rats were significantly elevated (P<0.01) on days 10, 20 and 30 PI, as well as the levels of TNF-α and INF-γ when compared to uninfected rodents. Regarding E-NTPDase activity in lymphocytes, it was possible to observe that the ATP hydrolysis was significantly decreased on days 20 (P<0.01) and 30 PI (P<0.05), while ADP hydrolysis was significantly reduced only on day 20 PI (P<0.01) when compared with uninfected group. Seric E-ADA activity had a significant reduction (P<0.01) during all three evaluated periods when compared to the control group, while E-ADA activity in lymphocytes increased significantly (P<0.01) when compared to the group A on day 10 PI; however on days 20 and 30 PI, its activity was considerable reduced in lymphocytes of infected animals (P<0.01). Therefore, it is possible to conclude that the infection caused by C. neoformans in immunocompetent rats leads to changes in the purinergic signaling (NTPDase and E-ADA), concomitantly with an inflammatory response (increased levels of cytokines and immunoglobulins) associated with inflammatory infiltrates and histological lesions in the lung.
Alves, Daniella de Sousa Mendes Moreira; Moraes, Aline Silva; Alves, Luciano Moreira; Gurgel-Gonçalves, Rodrigo; Lino Junior, Ruy de Souza; Cuba-Cuba, César Augusto; Vinaud, Marina Clare
T4 is the Acanthamoeba genotype most related to cases of granulomatous amoebic encephalitis (GAE) in immunocompromised patients and of keratitis in contact lens wearers. The determination of the pathogenic potential of Acanthamoeba clinical and environmental isolates using experimental models is extremely important to elucidate the capacity of free-living organisms to establish and cause disease in hosts. The aim of this study was to compare and evaluate the histopathology and culture between two different routes of experimental infection of T4 Acanthamoeba isolated from environmental and clinical source in mice (intracranial and intraperitoneal). Swiss isogenic healthy mice were inoculated with 10(4) trophozoites by intracranial (IC) and intraperitoneal (IP) routes and observed during 21 days. The brains from animals inoculated by the IC route were collected and from the animals of the IP inoculation group, the brains, livers, kidneys, spleens, and lungs were removed. The organs were prepared and appropriately divided to be evaluated with histopathology and culture. There was no significant difference between the inoculation routes in terms of isolates recovery (χ(2) = 0.09; p = 0.76). In the IC group, isolate recovery rate was significantly higher in histopathology than the one achieved by culture (χ(2) = 6.45; p < 0.01). Experimental infection revealed that all isolates inoculated could be considered invasive because it was possible to recover evolutive forms of Acanthamoeba in both routes. This work represents the first in vivo pathogenicity assay of primary isolation source in Central region of Brazil showing in vivo pathogenicity and hematogenous spread capacity of these protozoa, improving the knowledge on free-living amoebae isolates.
Bateman, K S; Munro, J; Uglow, B; Small, H J; Stentiford, G D
White spot syndrome virus (WSSV) is the most important pathogen known to affect the sustainability and growth of the global penaeid shrimp farming industry. Although most commonly associated with penaeid shrimp farmed in warm waters, WSSV is also able to infect, cause disease in and kill a wide range of other decapod crustaceans, including lobsters, from temperate regions. In 2005, the European Union imported US$500 million worth of raw frozen or cooked frozen commodity products, much of which originated in regions positive for white spot disease (WSD). The presence of WSSV within the UK food market was verified by means of nested PCR performed on samples collected from a small-scale survey of supermarket commodity shrimp. Passage trials using inoculum derived from commodity shrimp from supermarkets and delivered by injection to specific pathogen-free Pacific white shrimp Litopenaeus vannamei led to rapid mortality and pathognomonic signs of WSD in the shrimp, demonstrating that WSSV present within commodity shrimp was viable. We exposed a representative European decapod crustacean, the European lobster Homarus gammarus, to a single feeding of WSSV-positive, supermarket-derived commodity shrimp, and to positive control material (L. vannamei infected with a high dose of WSSV). These trials demonstrated that lobsters fed positive control (high dose) frozen raw products succumbed to WSD and displayed pathognomonic signs associated with the disease as determined by means of histology and transmission electron microscopy. Lobsters fed WSSV-positive, supermarket-derived commodity shrimp (low dose) did not succumb to WSD (no mortality or pathognomonic signs of WSD) but demonstrated a low level or latent infection via PCR. This study confirms susceptibility of H. gammarus to WSSV via single feedings of previously frozen raw shrimp products obtained directly from supermarkets.
Horn, J; Schlegel, U; Krettek, C; Ito, K
Infection continues to be a problematic complication of fracture treatment, particularly in severe open fractures. The implant design and material as well as implantation technique play an important role in the pathogenesis of local infection. The aim of our study was to determine if the local resistance to infection of a cannulated IM nail is less than that of a solid nail and more similar to that of a hollow nail. In 65 female White Zealand rabbits, the intramedullary cavity was inoculated with matching concentrations of Staphylococcus aureus, and one of the three nails was inserted. The solid nail had a greater than twofold higher resistance to infection (23%) compared to that of the other two nails (hollow, 65%; cannulated, 61%) which was statistically significant (p<0.02). No difference in infection resistance was detected between the hollow slotted and cannulated nail designs (p=1). Although these experimental results may be clinically considered, direct extrapolation to clinical infection rates is ill advised.
Chaudhari, Atul A; Kariyawasam, Subhashinie
The present study describes an experimental infection model for avian pathogenic Escherichia coli (APEC)-induced egg peritonitis in layer chickens. First, a pilot study which consisted of two separate experiments was carried out to compare two routes of inoculations of APEC to induce peritonitis and to examine if the presence of egg yolk in the peritoneum would facilitate APEC-induced peritonitis. This study showed that the presence of egg yolk in the peritoneum facilitated the development of egg peritonitis when the APEC was inoculated via the intra-uterine (IU) route. Based on the results of the pilot study, 56-wk-old white leghorn hens were divided into two groups of five chickens, Group G (inoculated with E. coli APECO78 strain) and Group H (control). Both groups were inoculated with 2-3 ml of egg yolk via the intraperitoneal route (IP). Subsequently, hens in Group H were inoculated with only egg yolk whereas the hens in Group G were inoculated with 1 x 10(9) colony-forming units of APECO78 bacteria via the IU route. Parameters such as mortality, clinical signs (anorexia, depression, and egg production efficiency), gross lesion scores, bacterial loads in internal organs, and histopathology of ovary and oviduct were assessed to evaluate the success of the infection model. Group G showed 40% acute mortality, severe depression, and anorexia with markedly reduced egg production and developed peritonitis-associated lesions such as accumulation of yellowish caseous fluid in the peritoneum, salpingitis, and oophoritis. Histopathologically, ovarian and oviduct tissues from group G exhibited severe inflammatory changes such as infiltration of mononuclear cells and edema. Group G also showed significant bacterial loads in the peritoneum, ovary, and oviduct. Interestingly, deceased birds from group G had also developed mild perihepatitis and pericarditis with heavy bacterial loads in the internal organs. On the other hand, group H birds did not exhibit any of the clinical
Lassen, Brian; Bangoura, Berit; Lepik, Triin; Orro, Toomas
Acute phase proteins (APPs) have been demonstrated to be useful in evaluating general health stress and diseases in cattle. Serum amyloid A (SAA) and haptoglobin (Hp) are APPs that are produced during inflammation, and likely play a role in host immunological defence against Eimeria infection and the associated intestinal tissue damage. We investigated the involvement of SAA and HP in an experimental study, including three groups of calves: a control group (group 0, n=11), and two groups infected with either 150,000 or 250,000 Eimeria zuernii oocysts (group 1 (n=11) and group 2 (n=12), respectively). The calves were monitored for 28 days and data was collected on oocyst excretion, faecal score, animal weight, and SAA and Hp serum concentrations. Generalized linear mixed models showed that the clinical symptoms, indicated by an increase in the number of oocysts in the faeces and severe diarrhoea, manifested at patency for group 1 and 2. Serum Hp and SAA levels also increased during this period. Hp appeared to be a more sensitive marker than SAA, and differences between groups 1 and 2 were observed only for Hp. Linear regression models showed a negative association between weight gain and Hp concentrations, calculated as the area under the curve (AUC) during the overall experimental period and the patency period. A similar result was seen for SAA only during the patency period. This result supports the assumption that reduced weight gain due to E. zuernii infection is an immunologically driven process that involves an increase in APPs. A random intercept regression model of oocyst shedding groups showed that calves shedding 1-500 oocysts had reduced concentrations of Hp, indicating that a different immunological reaction occurs during mild shedding of E. zuernii oocysts than during more intensive shedding. A similar model was used to examine associations between faecal scores and Hp concentrations for each group. Group 2 calves with haemorrhagic diarrhoea displayed
Shen, Chanjuan; Cheng, Anchun; Wang, Mingshu; Xu, Chao; Jia, Renyong; Chen, Xiaoyue; Zhu, Dekang; Luo, Qihui; Cui, Hengmin; Zhou, Yi; Wang, Yin; Xu, Zhiwen; Chen, Zhengli; Wang, Xiaoyu
To determine the expression and distribution of tegument proteins encoded by duck enteritis virus (DEV) UL51 gene in tissues of experimentally infected ducks, for the first time, an immunoperoxidase staining method to detect UL51 protein (UL51p) in paraffin-embedded tissues is reported. A rabbit anti-UL51 polyclonal serum, raised against a recombinant 6-His-UL51 fusion protein expressed in Escherichia coli, was prepared, purified, and used as primary antibodies. Fifty-eight 30-day-old DEV-free ducks were intramuscularly inoculated with the pathogenic DEV CHv strain as infection group, and two ducks were selected as preinfection group. The tissues were collected at sequential time points between 2 and 480 hr postinoculation (PI) and prepared for immunoperoxidase staining. DEV UL51p was first found in the spleen and liver at 8 hr PI; in the bursa of Fabricius and thymus at 12 hr PI; in the Harders glands, esophagus, small intestine (including the duodenum, jejunum, and ileum), and large intestine (including the caecum and rectum) at 24 hr PI; in the glandularis ventriculus at 48 hr PI; and in the pancreas, cerebrum, kidney, lung, and myocardium at 72 hr PI. Throughout the infection process, the UL51p was not seen in the muscle. Furthermore, the intensity of positive staining of DEV UL51p antigen in various tissues increased sharply from 8 to 96 hr PI, peaked during 120-144 hr PI, and then decreased steadily from 216 to 480 hr PI, suggesting that the expressional levels of DEV UL51p in systemic organs have a close correlation with the progression of duck virus enteritis (DVE) disease. A number of DEV UL51p was distributed in the bursa of Fabricius, thymus, spleen, liver, esophagus, small intestine, and large intestine of DEV-infected ducks, whereas less DEV UL51p was distributed in the Harders glands, glandularis ventriculus, cerebrum, kidney, lung, pancreas, and myocardium of DEV-infected ducks. Moreover, DEV UL51p can be expressed in the cytoplasm of various types
Moura, Vania B L; Silva, Mayara M; Batista, Lucas F; Gomes, Clayson M; Leenen, Pieter J M; Lino, Ruy S; Oliveira, Milton A P
Murine infection with Taenia crassiceps cysticerci is used as an experimental model for human and animal cysticercosis. In this infection parasites can be found associated with an inflammatory infiltrate enriched with macrophages. Experimental evidence exists supporting a role for either NO-producing classically activated (CAMΦ) or arginase- and CD301-expressing alternatively activated macrophages (AAMΦ) in T. crassiceps resistance. In both cell types, arginine is utilized as an important mediator in macrophage effector functions. To investigate whether there is an association between arginine availability, susceptibility to T. crassiceps and other parameters such as fibrosis, BALB/c mice were infected intraperitoneally with cysticerci and treated daily with the arginase inhibitor nor-NOHA or supplemented with l-arginine and followed for eight weeks. The numbers and developmental stages of parasites were evaluated as well as the presence of CD301+ AAMΦ, arginase activity and collagen deposition in the peritoneal membrane. Treatment with the arginase inhibitor or supplementation with l-arginine did not change the parasitic load or profile of the infection. However, the arginase inhibitor significantly decreased the deposition of collagen. These results suggest that arginase activity does not interfere with parasite control during experimental infection with T. crassiceps, but it is important for fibrosis in cysticercosis.
Hutton, R D; Kerbs, S; Yee, K
Trichophyton mentagrophytes invasion of guinea pig skin was examined by scanning electron microscopy. Biopsies were obtained daily for 12 days from experimental infection sites. Dermatophyte invasion, examined in detail by scanning electron microscopy of cross-sectioned, prefixed skin was evidenced by: the appearance of hyphae within the stratum corneum; follicular invasion by hyphae, which remained initially within the follicle wall; emergence of the hyphae from the wall into the follicular canal; proliferation of the fungus down the follicle, with furrowing of the follicle wall and hair shaft cuticle; penetration of hyphae into the hair shaft by subcuticular and transcuticular routes; and massive peripilar hyphal proliferation with arthrosporogenesis. A three-dimensional perception of the invasion sequence of a dermatophyte in guinea pig skin was obtained by scanning electron microscopy. Images PMID:711318
Kia, Eshrat Beigom
Echinococcus multilocularis, the small fox tapeworm, has an extensive geographical range in the northern hemisphere where foxes and small rodents represent natural hosts. The larval stage of this parasite, alveolar echinococcosis (AE), is an emerging zoonosis of increasing importance. It is a serious human illness which is often misdiagnosed as hepatic cancer. If not identified at an early stage of parasite development it can lead to the death of patients. Histological examination of biopsies is one of the classical methods of diagnosis. In this study, in order to gain unequivocal histopathological diagnosis of AE, the immunoperoxidase staining technique was performed on routinely processed histological sections of an experimentally infected gerbil, using rabbit anti-E. multilocularis protoscolex IgG labelled with horseradish peroxidase. Demonstration of AE antigen was achieved by dark brown stain of cyst membranes against a blue background of the host liver cells stained with hematoxylin.
de Kraker, Marlieke E A; Wolkewitz, Martin; Davey, Peter G; Koller, Walter; Berger, Jutta; Nagler, Jan; Icket, Claudine; Kalenic, Smilja; Horvatic, Jasminka; Seifert, Harald; Kaasch, Achim J; Paniara, Olga; Argyropoulou, Athina; Bompola, Maria; Smyth, Edmond; Skally, Mairead; Raglio, Annibale; Dumpis, Uga; Kelmere, Agita Melbarde; Borg, Michael; Xuereb, Deborah; Ghita, Mihaela C; Noble, Michelle; Kolman, Jana; Grabljevec, Stanko; Turner, David; Lansbury, Louise; Grundmann, Hajo
Antimicrobial resistance is threatening the successful management of nosocomial infections worldwide. Despite the therapeutic limitations imposed by methicillin-resistant Staphylococcus aureus (MRSA), its clinical impact is still debated. The objective of this study was to estimate the excess mortality and length of hospital stay (LOS) associated with MRSA bloodstream infections (BSI) in European hospitals. Between July 2007 and June 2008, a multicenter, prospective, parallel matched-cohort study was carried out in 13 tertiary care hospitals in as many European countries. Cohort I consisted of patients with MRSA BSI and cohort II of patients with methicillin-susceptible S. aureus (MSSA) BSI. The patients in both cohorts were matched for LOS prior to the onset of BSI with patients free of the respective BSI. Cohort I consisted of 248 MRSA patients and 453 controls and cohort II of 618 MSSA patients and 1,170 controls. Compared to the controls, MRSA patients had higher 30-day mortality (adjusted odds ratio [aOR] = 4.4) and higher hospital mortality (adjusted hazard ratio [aHR] = 3.5). Their excess LOS was 9.2 days. MSSA patients also had higher 30-day (aOR = 2.4) and hospital (aHR = 3.1) mortality and an excess LOS of 8.6 days. When the outcomes from the two cohorts were compared, an effect attributable to methicillin resistance was found for 30-day mortality (OR = 1.8; P = 0.04), but not for hospital mortality (HR = 1.1; P = 0.63) or LOS (difference = 0.6 days; P = 0.96). Irrespective of methicillin susceptibility, S. aureus BSI has a significant impact on morbidity and mortality. In addition, MRSA BSI leads to a fatal outcome more frequently than MSSA BSI. Infection control efforts in hospitals should aim to contain infections caused by both resistant and susceptible S. aureus.
Teixeira, Clarissa; Carvalho, Augusto M.; Mattos, Paulo S.; Cristal, Juqueline R.; Muniz, Aline C.; Miranda, José Carlos; Barral, Aldina
Background Previous works showed that immunization with saliva from Lutzomyia intermedia, a vector of Leishmania braziliensis, does not protect against experimental infection. However, L. braziliensis is also transmitted by Lutzomyia whitmani, a sand fly species closely related to Lu. intermedia. Herein we describe the immune response following immunization with Lu. whitmani saliva and the outcome of this response after L. braziliensis infection. Methods and findings BALB/c mice immunized with Lu. whitmani saliva developed robust humoral and cellular immune responses, the latter characterized by an intense cellular infiltrate and production of IFN-γ and IL-10, by both CD4+ and CD8+ cells. Mice immunized as above and challenged with L. braziliensis plus Lu. whitmani saliva displayed significantly smaller lesions and parasite load at the challenge site. This protection was associated with a higher (p<0.05) IFN-γ production in response to SLA stimulation. Long-term persisting immunity was also detected in mice immunized with Lu. whitmani saliva. Furthermore, individuals residing in an endemic area for cutaneous leishmaniasis (CL) presented antibody responses to Lu. whitmani saliva. However CL patients, with active lesions, displayed a lower humoral response to Lu. whitmani saliva compared to individuals with subclinical Leishmania infection. Conclusion Pre-exposure to Lu. whitmani saliva induces protection against L. braziliensis in a murine model. We also show that Lu. whitmani salivary proteins are immunogenic in naturally exposed individuals. Our results reinforce the importance of investigating the immunomodulatory effect of saliva from different species of closely related sand flies. PMID:27812113
Rhyan, Jack; McCollum, Matthew; Gidlewski, Thomas; Shalev, Moshe; Ward, Gordon; Donahue, Brenda; Arzt, Jonathan; Stenfeldt, Carolina; Mohamed, Fawzi; Nol, Pauline; Deng, Ming; Metwally, Samia; McKenna, Thomas; Salman, Mo
There is limited information on the pathogenesis and epidemiology of foot-and-mouth disease (FMD) in North American wildlife and none concerning pronghorn ( Antilocapra americana ). In an experimental study of 13 pronghorn and six steers ( Bos taurus ), we compared the susceptibility of pronghorn to FMD virus (FMDV) strain O, with that of cattle ( Bos taurus ). We also determined the potential for intra- and interspecies transmission of FMDV strain O in pronghorn and cattle, assessed the application of conventional laboratory tests in their suitability to detect FMDV infection in pronghorn, and evaluated the potential role of pronghorn as efficient long-term carriers of FMDV. After acclimation to containment at Plum Island Animal Disease Center, two pronghorn and one steer were each infected by intraepithelial tongue inoculation with 10,000 bovine tongue infective doses of FMDV, strain O1 Manisa. Inoculated animals were housed with contact animals. When contact-exposed animals developed fever they were placed in rooms with previously unexposed animals. All inoculated and exposed cattle and pronghorn developed clinical disease typical of FMD. Pronghorn developed severe foot lesions and mild to moderate oral lesions, primarily on the tongue. Duration of clinical signs in both species was 2-3 wk with foot abnormalities evident to the end of the study (51 d postexposure). Other lesions included pancreatitis, myositis of the tongue, and secondary lesions including pleuritis, pneumonia, decubital ulcers, and tenosynovitis. Virus transmission occurred between pronghorn, from cattle to pronghorn, and from pronghorn to cattle. Conventional laboratory tests detected virus and antibodies against nonstructural and structural FMDV proteins in pronghorn and cattle. Virus was present in some animals for 1 wk but was not detectable by virus isolation or PCR at 3 wk postinfection or afterward.
Xiao, Xia; Sun, Jian; Yang, Tao; Fang, Xi; Cheng, Jie; Xiong, Yan Q.; Liu, Ya-Hong
Mycoplasma gallisepticum is the most important pathogen in poultry among four pathogenic Mycoplasma species. Tiamulin is a pleuromutilin antibiotic that shows a great activity against M. gallisepticum and has been approved for use in veterinary medicine particularly for poultry. However, the pharmacokinetic/pharmacodynamics (PK/PD) profiles of tiamulin against M. gallisepticum are not well understood. Therefore, in the current studies, we investigated the in vivo PK/PD profiles of tiamulin using a well-established experimental intratracheal infection model of M. gallisepticum. The efficacy of tiamulin against M. gallisepticum was studied in 8-day-old chickens after intramuscular (i.m.) administration at 10 doses between 0–80 mg/kg. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to evaluate the PK parameters of tiamulin following i.m. administration at doses of 5, 40, and 80 mg/kg in Mycoplasma gallisepticum-infected neutropenic chickens. Real-time PCR (RT-PCR) was used for quantitative detection of M. gallisepticum. The MIC of tiamulin against M. gallisepticum strain S6 was 0.03 μg/mL. The PK/PD index, AUC24h/MIC, correlated well with the in vivo antibacterial efficacy. The in vivo data suggest that animal dosage regimens should supply AUC24h/MIC of tiamulin of 382.68 h for 2 log10 ccu equivalents M. gallisepticum reduction. To attain that goal, the administered dose is expected to be 45 mg/kg b.w. for treatment of M. gallisepticum infection with an MIC90 of 0.03 μg/mL. PMID:27656647
Banerjee, Soumya; Guedj, Jeremie; Ribeiro, Ruy M; Moses, Melanie; Perelson, Alan S
West Nile virus (WNV) is an emerging pathogen that has decimated bird populations and caused severe outbreaks of viral encephalitis in humans. Currently, little is known about the within-host viral kinetics of WNV during infection. We developed mathematical models to describe viral replication, spread and host immune response in wild-type and immunocompromised mice. Our approach fits a target cell-limited model to viremia data from immunocompromised knockout mice and an adaptive immune response model to data from wild-type mice. Using this approach, we first estimate parameters governing viral production and viral spread in the host using simple models without immune responses. We then use these parameters in a more complex immune response model to characterize the dynamics of the humoral immune response. Despite substantial uncertainty in input parameters, our analysis generates relatively precise estimates of important viral characteristics that are composed of nonlinear combinations of model parameters: we estimate the mean within-host basic reproductive number,R0, to be 2.3 (95% of values in the range 1.7-2.9); the mean infectious virion burst size to be 2.9 plaque-forming units (95% of values in the range 1.7-4.7); and the average number of cells infected per infectious virion to be between 0.3 and 0.99. Our analysis gives mechanistic insights into the dynamics of WNV infection and produces estimates of viral characteristics that are difficult to measure experimentally. These models are a first step towards a quantitative understanding of the timing and effectiveness of the humoral immune response in reducing host viremia and consequently the epidemic spread of WNV.
Hurtenbach, U; Böggemeyer, E; Stehle, T; Museteanu, C; Del Pozo, E; Simon, M M
Glucocorticosteroids (GC) are widely used as anti-inflammatory agents. The effects of Prednisolone on the development of Borrelia (B.) burgdorferi-induced clinical arthritis and organ inflammation was studied in severe combined immunodeficiency (SCID) mice. The drug was administered orally at a dose of 3, 10 and 30 mg/kg, starting shortly before experimental infection of the mice. A dose dependent inhibition of arthritic joint swelling was observed. Full protection was obtained with 30 mg/kg until 21 days after infection, subsequently, mild joint swelling developed but progression and severity of the disease was considerably less than in the other treated as well as in the untreated mice. Inhibition of clinical arthritis coincided with reduction of inflammatory cell infiltration in the joints, liver and muscle. Prednisolone was ineffective when application was initiated after arthritis was fully developed, i.e., 22 days after infection. Since the activated endothelium plays a critical role in development of inflammatory lesions, the expression of the cellular adhesion molecules (CAMs) E-selectin, P-selectin, ICAM-1 and VCAM-1 was determined in vitro using the bEnd3 endothelial cell line. Stimulation with a sonicated B. burgdorferi preparation in the presence of the water-soluble compound Prednisolone-21-hemisuccinate considerably reduced expression of ICAM-1, and marginally also of E-selectin, whereas the level of P-selectin and VCAM-1 remained unaltered. Thus, downregulation of ICAM-1 might be a critical factor in Prednisolone-mediated inhibition of B. burgdorferi-induced inflammation; the flare up of the disease after the initial protection indicates that additional therapy, e.g. with antibiotics, is necessary.
Taus, Naomi S; O'Toole, Donal; Herndon, David R; Cunha, Cristina W; Warg, Janet V; Seal, Bruce S; Brooking, Angela; Li, Hong
Malignant catarrhal fever (MCF), due to ovine herpesvirus 2 (OvHV-2), causes appreciable death loss in ranched bison (Bison bison) throughout North America. No vaccine exists to protect animals from disease. Since OvHV-2 has not been propagated in vitro, one strategy to develop a modified live vaccine is to use a closely related, non-pathogenic member of the malignant catarrhal fever virus family as a vector expressing potentially protective OvHV-2 epitopes. To date, no controlled experimental challenge studies with alcelaphine herpesvirus 2 (AlHV-2) derived from topi (Damaliscus lunatus jimela) have been reported The unique or light DNA segment of the AlHV-2 genome was sequenced and annotated and the virus was tested for its ability to infect and induce disease in American bison. Yearling bison were inoculated intranasally (n=4) or intramuscularly (n=3) with 2 × 10(-4.7) TCID50 of AlHV-2, and monitored for infection and the development of disease. Six inoculated bison became infected with AlHV-2. Two of the six animals developed clinical signs and had gross and histological lesions consistent with terminal MCF, which differed in distribution from those in bison with MCF due to OvHV-2. One other animal developed minor clinical signs and had gross and histological pulmonary lesions consistent with early (pre-clinical) stages of MCF. Unmodified low cell culture passage AlHV-2 derived from topi is an unsuitable vaccine vector for the prevention of MCF. However, the annotated genome might be useful in identifying genes which could be deleted to potentially attenuate the virus for bison.
Khalil, R M; Hültner, L; Mailhammer, R; Luz, A; Moeller, J; Mohamed, A A; Omran, S; Dörmer, P
It has been reported that interleukin-6 (IL-6) is expressed in cells of acute inflammatory granulomas experimentally induced in mice by eggs of Schistosoma mansoni. Moreover, in vitro IL-6 was shown to enhance the cytotoxic activity of human platelets against larvae of S. mansoni. To elucidate further a proposed biological significance of this cytokine during the course of schistosomiasis, we studied the kinetics of IL-6 production and concomitantly performed a histopathological analysis of the livers in BALB/c mice subcutaneously infected with S. mansoni cercariae. Over a period of 24 weeks postinfection (p.i.) we monitored serum IL-6 levels, IL-6 production in vitro by pokeweed mitogen (PWM)-stimulated spleen cells as well as IL-6 mRNA expression in livers, spleens and kidneys. We found significantly elevated IL-6 levels in PWM-stimulated spleen cell-conditioned media (SCM) at weeks 6 to 20 p.i., peaking at week 10 p.i. In contrast, serum IL-6 concentrations started to rise not before week 8 but remained significantly elevated above normal control values until week 24 p.i. The time pattern of enhanced IL-6 mRNA expression detected in spleens and livers, but not in kidneys, as well as the rises of IL-6 in SCM and with a delay of 2 weeks in serum samples correlated with the onset of the egg-induced inflammatory reactions as well as the incidence and the number of the granulomas observed histopathologically in the livers of infected mice. Our data emphasize both a local and a systemic role of IL-6 in the host immune response following infection of mice with S. mansoni. Images Figure 3 PMID:8943723
Tanideh, Nader; Rokhsari, Pedram; Mehrabani, Davood; Mohammadi Samani, Soleiman; Sabet Sarvestani, Fatemeh; Ashraf, Mohammad Javad; Koohi Hosseinabadi, Omid; Shamsian, Shahram; Ahmadi, Nasrollah
BACKGROUND Burn is still one of the most devastating injuries in emergency medicine while improvements in wound healing knowledge and technology have resulted into development of new dressings. This study was undertaken to evaluate the healing effect of licorice in Pseudomonas aeruginosa infected burn wounds of experimental rat model. METHODS One hundred and twenty female Sprague-Dawley rats were randomly allocated to 4 equal groups. Group A received silver sulfadiazine ointment, Group B received 10% licorice extract and Group C was considered as control group and received gel base as the base of medication. Group D did not receive any medication and just underwent burn injury. A standard 3rd degree burn wound was produced by a hot plate with similar size about 20% of total body surface area (TBSA) and at identical temperature. After 24 h of burn production, 108 colony forming units (CFU) of toxigenic strains of P. aeruginosa (PA 103) were inoculated subcutaneously into the burnt area. After 3, 7, 14, 21 and 28 days of therapy, the animals were sacrificed and burn areas were macroscopically examined and histologically evaluated. RESULTS Decrease in size of the burn wounds, in inflammation and re-epithelialization were poor in groups B-D. Infection to P. aeruginosa was still visible in groups B-D but was absent in Group A. The mean histological score, tensile strength, maximum stress, yield strength and stiffness in groups B-D were lower compared with Group A. CONCLUSION Licorice extract in 10% concentration was shown not to be effective in healing of P. aeruginosa infected burn wounds. PMID:25489532
Gast, Richard K; Guraya, Rupa; Jones, Deana R; Anderson, Kenneth E
The majority of human illnesses caused by Salmonella Enteritidis are attributed to contaminated eggs, and the prevalence of this pathogen in commercial laying flocks has been identified as a leading epidemiologic risk factor. Flock housing and management systems can affect opportunities for the introduction, transmission, and persistence of foodborne pathogens in poultry. The animal welfare implications of different types of housing for laying hens have been widely discussed in recent years, but the food safety consequences of these production systems remain incompletely understood. The present study assessed the effects of 2 different housing systems (conventional cages and colony cages enriched with perching and nesting areas) on the horizontal transmission of experimentally introduced Salmonella Enteritidis infection within groups of laying hens. In each of 2 trials, 136 hens were distributed among cages of both housing systems and approximately one-third of the hens in each cage were orally inoculated with doses of 10(8) cfu of Salmonella Enteritidis (phage type 13a in one trial and phage type 4 in the other). At regular intervals through 23 d postinoculation, cloacal swabs were collected from all hens (inoculated and uninoculated) and cultured for Salmonella Enteritidis. Horizontal contact transmission of infection was observed for both Salmonella Enteritidis strains, reaching peak prevalence values of 27.1% of uninoculated hens in conventional cages and 22.7% in enriched cages. However, no significant differences (P > 0.05) in the overall frequencies of horizontal Salmonella Enteritidis transmission were evident between the 2 types of housing. These results suggest that opportunities for Salmonella Enteritidis infection to spread horizontally throughout laying flocks may be similar in conventional and enriched cage-based production systems.
Kanitz, Fábio A; Cargnelutti, Juliana F; Anziliero, Deniz; Gonçalves, Kelley V; Masuda, Eduardo K; Weiblen, Rudi; Flores, Eduardo F
Equid herpesvirus type 1 (EHV-1) is an important pathogen of horses worldwide, associated with respiratory, reproductive and/or neurological disease. A mouse model for EHV-1 infection has been established but fails to reproduce some important aspects of the viral pathogenesis. Then, we investigated the susceptibility of rabbits to EHV-1 aiming at proposing this species as an alternative model for EHV-1 infection. Weanling rabbits inoculated intranasal with EHV-1 Kentucky D (