Effects of dicopper oxide and copper sulfate on growth performance and gut microbiota in broilers.

An experiment was conducted to determine the effects of two sources of copper (Cu) from copper sulfate (CuSO4) and dicopper oxide (Cu2O, CoRouge) at three levels of inclusion (15, 75, and 150 mg/kg) on growth performance and gut microbiota of broilers. A total of 840 one-d-old male chickens (Ross 308) were weighed and randomly allocated to seven dietary treatments: negative control (NC, a basal diet without Cu addition), and the NC supplemented with 15, 75, or 150 mg Cu/kg from CuSO4 or Cu2O (12 replicate pens/treatment, 10 chicks per pen). Broilers were challenged by reusing an old litter with high concentrations in Clostridium perfringens to promote necrotic enteritis. Broiler performance was registered at d 21, 35, and 42. Excreta samples were collected at d 14, 28, and 42 for antimicrobial resistance (AMR) analyses. At d 43, one broiler per pen was euthanized to obtain ileal content for microbial characterization. Body weight d 35 and daily gain d 42 improved (P < 0.05) in Cu2O as Cu dose inclusion increased from 15 mg/kg to 150 mg/kg. Supplementation of 150 mg/kg of Cu from Cu2O decreased the abundance (P < 0.01) of some families such as Streptococcaceae and Corynebacteriaceae and increased the abundance (P < 0.05) of some commensal bacteria like Clostridiaceae and Peptostreptococcaceae. Phenotypic AMR was not different among treatments on d 14 and 28. Isolated Enterococcus spp. from broilers fed the NC diet on d 42 showed higher (P < 0.05) resistance to enrofloxacin, gentamicin, and chloramphenicol compared with Cu treatments. By contrast, the isolated Escherichia coli from broilers fed 150 mg/kg of Cu, either from CuSO4 or Cu2O, showed higher (P < 0.05) resistance to streptomycin and chloramphenicol compared to the NC. This study suggests that supplementing 150 mg/kg of Cu from Cu2O establishes changes in the gut microbiota by regulating the bacterial population in the ileum, which may explain the positive impact on broilers' growth performance.

Uterine serpin (SERPINA 14) correlates negatively with cytokine production at the foetal-maternal interface but not in the corpus luteum in pregnant dairy heifers experimentally infected with Neospora caninum.

This study examines gene expression patterns in dairy heifers experimentally infected with N. caninum during on Day 110 of pregnancy with live foetuses at euthanasia, 42 days later. The study population was constituted of four non-infected controls and three infected dams. Gene expression was determined on gamma interferon (IFNγ), (Th1 pro-inflammatory cytokine), interleukin-4 (IL4) (Th2 pro-gestation cytokine) or interleukin-10 (IL10) (T regulatory cytokine) and the serine peptidase inhibitor SERPINA14 in intercaruncular, placental, uterine lymph node (UTLN) and luteal tissue samples. Intercaruncular SERPINA14 expression was negatively correlated with IFNγ expression in cotyledon samples and with IL4 expression in UTLN. No relationships were detected between cytokine gene expression at the foetal-maternal interface and SERPINA14 expression in the luteal samples. Our findings suggest that gene expression of the uterine serpin SERPINA14 correlates negatively with the expression of Th1 and Th2 cytokines at the foetal-maternal interface but not in the corpus luteum.

Transmission of Porcine reproductive and respiratory syndrome virus 1 to and from vaccinated pigs in a one-to-one model.

The present study examined transmission by contact of Porcine reproductive and respiratory syndrome virus (PRRSV) 1 in a one-to-one model to vaccinated and unvaccinated pigs and from vaccinated infected pigs to other vaccinated pigs. The experiment started by randomly assigning weaned pigs to groups V (n=24) and U (n=26). V pigs were vaccinated with a commercial live attenuated PRRSV vaccine and the U animals were kept as unvaccinated controls. Twenty-eight days later, 6U pigs were separated and allocated in individual boxes. The remaining 20U pigs were intranasally inoculated with PRRSV isolate 3267 (from now on designated as seeder (S) pigs) and 48h later were distributed in boxes where they were commingled with either V or U pigs in 1:1 groups (first contact phase), resulting in 6S:U and 14S:V pairs. As soon as a V pig was detected to be viremic because of contact with a S, the infected V (from now on designated as Vinf) was transferred (<24h after detection) to a new pen where it was comingled with a new V pig (designated as V2) in a second contact phase. For the first contact phase, pigs were maintained 21days at maximum and for the second contact phase the maximum exposure period was 14days. Two V pigs tested positive for the vaccine virus (>99.5% similarity) when they were relocated with the corresponding V2 pigs and they were removed; thus, only 12Vinf were finally considered. All V pigs (12/12) exposed to S animals became infected although the first detection of viremia occurred at 13.6±3.6days, one week later than in U (p<0.05). Also, duration of viremia was shorter for Vinf compared to U, (5.5±4.3days versus 12.5±2.7days). The Vinf group showed remarkable individual variability: eight animals had a viremic period of 5 or less days (3.0±1.4) while the remaining four had a longer viremic period of more than one week (10.8±2.9). This situation was not observed in U. In the second contact phase, transmission from Vinf to V2 pigs occurred in 7/8 cases (87.5%). The mean duration of viremia for V2 was 4.8±3.4 and two different patterns were again observed: two animals had viremias of 9-10days and the rest averaged 3.0±1.4days (range: 2-5days). Vaccinated groups Vinf and V2 had a significantly lower PRRSV shedding in oral fluids for at least the first 9days after the onset of the viremia compared to U, and shedding for V2 was even significantly lower (p<0.05) than shedding for Vinf. Our experimental design reproduced the worst-case scenario for evaluating the effect of vaccination and, under such conditions; it was still efficacious in slowering PRRSV transmission and decreasing the global viral load and particularly oral shedding.

Cytokine gene expression in aborting and non-aborting dams and in their foetuses after experimental infection with Neospora caninum at 110 days of gestation.

Neospora caninum is a major cause of abortion in cattle. However, it is not known why not all infected animals abort. In this study, Th1 (IFN-γ), Th2 (IL4) and T reg (IL-10) cytokine gene expression was examined by real time PCR using the TaqMan approach in all of these dams and their foetuses after experimental infection with the isolate Nc-Spain7 at 110 days of pregnancy and euthanasia 6 weeks after infection. In prior published work, foetal death was observed in three of six infected dams and transplacental infection in all the 6 infected foetuses. In the spleen of the dams, IL-4 expression was down-regulated in dams with aborted/non viable foetuses compared to both uninfected dams (controls, n=3) and infected dams with live fetuses at euthanasia. In the lymph nodes draining the placenta, up-regulated expression of IL-4 was observed in infected dams with live foetuses compared to control dams. In the placenta, infected dams with live foetuses had significantly up-regulated IFN-γ in both caruncle and cotyledon and up-regulated IL-10 in cotyledon compared to control dams. Infected live foetuses showed up-regulated expression of IFN-γ and IL-10 in foetal spleen, and showed downregulated expression of IL-4 in the thymus compared to control uninfected foetuses. Expression of any cytokine in the thymus was significantly lower compared to the levels observed in foetal spleen. The results indicate an up-regulated expression of Th1, Th2 and Treg in infected dams with live foetuses and in their foetuses. On the other hand, down-regulation of Th2 immune responses and Treg cytokines were observed in infected dams which had aborted or had non-viable foetuses at euthanasia, suggesting an immunological recovery of cytokine gene expression levels in dams a few weeks after an abortion occurred.

Maternal and foetal cytokine production in dams naturally and experimentally infected with Neospora caninum on gestation day 110.

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.

Experimental Neospora Caninum Infection in Pregnant Dairy Heifers Raises Concentrations of Pregnancy-Associated Glycoproteins 1 and 2 in Foetal Fluids.

Plasma concentrations of PAG-1 are used for pregnancy diagnosis and as a marker of placental/foetal well-being, while those of PAG-2 may be an indicator of abortion risk in Neospora caninum-infected cows. Studies have shown that N. caninum infection modifies PAG-1 and PAG-2 patterns in maternal blood plasma. However, no prior work has examined the effects of N. caninum infection on concentrations of PAGs in foetal fluids. In this study, PAG-1, PAG-2 and pH levels were determined in the amniotic and allantoic fluids of foetuses collected at 152 days of gestation from control uninfected dams and from dams experimentally infected with N. caninum on Day 110 of gestation. Foetal fluids from infected foetuses had significantly higher PAG-2 concentrations (p = 0.026) and pH values (p = 0.02) than fluids from non-infected foetuses. In infected foetuses, significantly higher concentrations of PAG-1 (p < 0.001) and PAG-2 (p < 0.001) were detected in fluid samples showing antibodies against N. caninum than those without antibodies. Moreover, pH values were significantly higher (p = 0.011) in foetal fluid samples with antibodies than in samples from non-infected foetuses. In conclusion, this is the first report on the effect of N. caninum infection on PAG levels in foetal fluids. Our results indicate that following the experimental infection of dams with N. caninum on Day 110 of gestation, foetal fluids collected from the infected foetuses of these dams featured higher PAG-1 and PAG-2 levels and pH values than fluids from non-infected controls, provided that the samples tested showed the presence of antibodies. The clinical implications of these findings are that following infection with N. caninum, most cows will experience some level of placental damage and that this injury correlates with foetal fluid PAG levels and pH.

Maternal and fetal immune response patterns in heifers experimentally infected with Neospora caninum in the second trimester of pregnancy--a descriptive study.

Fetal and maternal immune responses 3, 6 and 9 weeks post infection (wpi) were investigated in cows experimentally infected with Neospora caninum on day 110 of gestation. Descriptive analysis showed that the fetuses had lower percentages of spleen T cell subpopulations (CD3+, CD4+ and CD8+) at 6 wpi compared to 3 wpi and/or 9 wpi, with the lowest percentages observed in a dead fetus found upon euthanasia at that time. Increased expression of most cytokines over levels recorded at 3 and 9 wpi were found in fetuses that were alive at 6 wpi. Up-regulated Th1, Th2 and Treg expression was also observed at 6 wpi in the spleen and in the lymph nodes draining the placenta of the cows. At the placental level, while most cytokines were down-regulated from 6 wpi, up-regulation of IL-4 expression was observed at 6 wpi in the caruncle. Our results suggest that the immune response at 6 wpi was crucial for fetal survival in this model of bovine neosporosis.

Oral human papillomavirus type-specific infection in HIV-infected men: a prospective cohort study among men who have sex with men and heterosexual men.

The natural history of type-specific oral infection of human papillomavirus (HPV) was assessed in a cohort of HIV-infected men (538 men who have sex with men (MSM); 195 heterosexuals). Risk factors associated with oral HPV infections were examined. The overall prevalence of HPV was 16%: HPV-16 was the most prevalent type (3.7% MSM; 7.8% heterosexuals). The prevalence of HPV-16 in heterosexuals was associated with CD4 nadir counts <200 cells/µL (ORadjusted = 3.0, 95% CI, 1.4-6.3). The overall incidence of HPV was similar between groups (11%), but the incidence of HPV-16 was higher in heterosexuals (ORadjusted = 3.2, 95% CI, 1.1-9.5). Not only MSM but also HIV-infected heterosexual men are at risk of HPV infection. Regular and careful oral inspection is needed.

Analysis of ORF5 and full-length genome sequences of porcine reproductive and respiratory syndrome virus isolates of genotypes 1 and 2 retrieved worldwide provides evidence that recombination is a common phenomenon and may produce mosaic isolates.

UNLABELLED: Recombination is currently recognized as a factor for high genetic diversity, but the frequency of such recombination events and the genome segments involved are not well known. In the present study, we initially focused on the detection of recombinant porcine reproductive and respiratory syndrome virus (PRRSV) isolates by examining previously published data sets of ORF5 sequences (genotypes 1 and 2) obtained worldwide. We then examined full-length genome sequences in order to determine potential recombination breakpoints along the viral genome. For ORF5, 11 sets of genotype 1 sequences from different geographical areas, including 2 Asian, 1 American, and 7 European regions, and three sets of genotype 2, including sets from China, Mexico, and the United States, were analyzed separately. Potential recombination breakpoints were detected in 10/11 genotype 1 sets, including 9 cases in which the clustering of at least one isolate was different before and after the breakpoints. In genotype 2, potential breakpoints and different tree clustering of at least one strain before and after the breakpoint were observed in 2 out of 3 sets. The results indicated that most of the ORF5 data sets contained at least one recombinant sequence. When the full-length genome sequences were examined, both genotype 1 and 2 sets presented breakpoints (10 and 9, respectively), resulting in significantly different topologies before and after the breakpoints. Mosaic genomes were detected in genotype 1 sequences. These results may have significant implications for the understanding of the molecular epidemiology of PRRSV. IMPORTANCE: PRRSV is one of the most important viruses affecting swine production worldwide, causing big economic losses and sanitary problems. One of the key questions on PRRSV arises from its genetic diversity, which is thought to have a direct impact on immunobiology, epidemiology, diagnosis, and vaccine efficacy. One of the causes of this genetic diversity is recombination among strains. This study provides evidence that recombinant PRRSV isolates are common in most of the countries with significant swine production, especially PRRSV genotype 1. This observation has implications in the proper characterization of PRRSV strains, in the future development of phylogenetic studies, and in the development of new PRRSV control strategies. Moreover, the present paper emphasizes the need for a deeper understanding of the mechanisms and circumstances involved in the generation of genetic diversity of PRRSV.

Fatal toxoplasmosis associated with an atypical Toxoplasma gondii strain in a Bennett's wallaby (Macropus rufogriseus) in Spain.

Toxoplasmosis is often fatal in captive wallabies, but the causes of this high susceptibility are not well understood. Here, we report fatal toxoplasmosis in a Bennet's wallaby (Macropus rufogriseus) due to an atypical Toxoplasma gondii strain for the first time in Europe. The wallaby was from a colony of 7 Bennet's wallabies that died over a 17-month period at a safari-zoological park in northeastern Spain. Only one of these wallabies was examined at necropsy. T. gondii-like organisms were detected by histological examination in several tissues and the diagnosis was confirmed through detection of T. gondii DNA by PCR. A nested PCR-based assay detected the 200- to 300-fold repetitive 529 bp DNA fragment of T. gondii in a sample of brain tissue. Genotyping analysis with 15 single-copy microsatellite markers was performed on this positive DNA sample and revealed an atypical genotype. Atypical genotypes are frequently associated with severe forms of toxoplasmosis in humans. The present report highlights the possible implications of the introduction of new atypical, more pathogenic T. gondii strains, to non-endemic areas.

Circumcision and penile human papillomavirus prevalence in human immunodeficiency virus-infected men: heterosexual and men who have sex with men.

Male circumcision is associated with a lower risk of penile human papillomavirus (HPV) infection in human immunodeficiency virus (HIV) uninfected men. Few studies have evaluated the role of male circumcision in penile HPV infection in HIV-infected men. The aim of this cross-sectional study was to examine the association between male circumcision and the prevalence of penile HPV infection among HIV-infected men-both men who have sex with men (MSM) and heterosexual men. Samples from 706 consecutive men included in the CARH-MEN cohort (overall 24% circumcised: 26% of MSM, 18% of heterosexual men) were examined by Multiplex-PCR. In the overall group (all HIV-infected men included), the prevalence of any penile HPV infection was 22% in circumcised men and 27% in uncircumcised men (OR = 1.0, 95% CI 0.6-1.6, adjusted analysis). In the circumcised group the overall prevalence of HPV infection was 22% in MSM and 24% in the heterosexual men, whereas in the uncircumcised group the prevalence was 26% and 28%, respectively. The prevalence of high-risk HPV types tended to be lower in the circumcised MSM (14% vs 21%, OR = 0.6, 95% CI 0.3-1.1, p 0.088), but it was similar in the heterosexual men (18% in circumcised vs 20% in uncircumcised). These results suggest that male circumcision may be associated with a lower prevalence of oncogenic high-risk penile HPV infection in HIV-infected MSM.

Condylomata, cytological abnormalities and human papillomavirus infection in the anal canal in HIV-infected men.

BACKGROUND: Genital infections with low-risk (LR) and high-risk (HR) human papillomavirus (HPV) genotypes are associated with ano-genital condylomata and anal squamous cell cancer. HPV-related pathologies in HIV-infected men are a serious concern. In this study, the prevalence of anal condylomata and their association with cytological abnormalities and HPV infection in the anal canal in HIV-infected men [men who have sex with men (MSM) and heterosexuals] were estimated. METHODS: This was a cross-sectional study based on the first visits of patients in the Can Ruti HIV-positive Men (CARH·MEN) cohort. Anal condylomata were assessed by clinical and proctological examination. Samples from the anal canal were collected for HPV genotyping and cytological diagnoses. RESULTS: A total of 640 HIV-infected men (473 MSM and 167 heterosexuals) were included in the study. The overall prevalence of anal condylomata was 25% [157 of 640; 95% confidence interval (CI) 21-28%]; in MSM it was 28% and in heterosexuals it was 15% [odds ratio (OR) 2.2; 95% CI 1.4-3.5]. In patients with anal condylomata, HPV infection in the anal canal was more prevalent (92% vs. 67% in those without anal condylomata; OR 8.5; 95% CI 3.2-22). This higher HPV prevalence involved at least two HPV genotypes (OR 4.0; 95% CI 2.2-7.1), mainly HR genotypes (OR 3.3; 95% CI 1.7-6.4). Similarly, the cumulative prevalence of HPV-6 and HPV-11 was higher in patients with anal condylomata (63% vs. 19% in those without anal condylomata). Having anal condylomata was associated with higher prevalences of cytological abnormalities (83% vs. 32% in those without anal condylomata; OR 6.9; 95% CI 3.8-12.7) and high-grade squamous intraepithelial lesions (HSILs) (9% vs. 3% in those without anal condylomata; OR 9.0; 95% CI 2.9-28.4) in the anal canal. CONCLUSIONS: HIV-infected men with anal condylomata were at risk of presenting HSILs and harbouring multiple HR HPV infections in the anal canal. Although MSM presented the highest prevalence of anal condylomata, heterosexual men also had a clinically important prevalence. Our findings emphasize the importance of screening and follow-up for condylomata in the anal canal in HIV-infected men.

Effects of challenge with a virulent genotype II strain of porcine reproductive and respiratory syndrome virus on piglets vaccinated with an attenuated genotype I strain vaccine.

Porcine reproductive and respiratory syndrome virus (PRRSV) is endemic in most parts of Asia, where genotype I and II strains of diverse virulence may coexist. This study evaluated the outcome of infection with a highly virulent Asian genotype II PRRSV isolate in piglets vaccinated with a genotype I vaccine. Twenty-one 3-week-old piglets were divided in three groups: Pigs in group V (n=8) were vaccinated with an attenuated genotype I commercial PRRSV vaccine, while pigs in group U (n=8) and a control group (group C; n=5) were unvaccinated; 6 weeks later, pigs in groups V and U were challenged intranasally with a highly virulent strain of genotype II PRRSV (1×10(5) 50% tissue culture infectious doses/mL), while pigs in group C received a placebo. Over a period of 21 days after challenge, vaccinated pigs had significantly lower mortality (0/8 versus 2/8), fewer days of fever, a lower frequency of catarrhal bronchopneumonia, higher weight gains (13.4 versus 6.6 kg) and lower levels of viraemia compared to unvaccinated challenged pigs. Immunisation with a genotype I attenuated PRRSV vaccine provided partial protection against challenge with a highly virulent genotype II strain.

Cytokine gene expression profiles in peripheral blood mononuclear cells from Neospora caninum naturally infected dams throughout gestation.

Neospora caninum is a major cause of abortion in cattle but it is not known why some infected animals suffer abortion while others do not. An essential role in protective immunity against N. caninum has been proposed for Th1 cytokines such as IFN-γ and IL-12 although cytokine patterns in N. caninum infected pregnant cattle have been scarcely addressed. In this study, gene expression of the cytokines IFN-γ, IL-12, IL-10, IL-4 and TNF-α was analyzed by real time RT-PCR in peripheral blood mononuclear cells in N. caninum naturally infected dams throughout pregnancy. Blood samples were drawn from 18 cows (13 N. caninum seropositive and 5 N. caninum seronegative) on Days 45, 90, 120, 150, 180 and 210 of pregnancy or until abortion. Four seropositive animals aborted. Compared to the seronegative animals, N. caninum infected dams showed up-regulated mRNA levels of the Th1 cytokines, IFN-γ, TNF-α and IL-12p40, along with up-regulation of the T regulatory (Treg) cytokine IL-10. In contrast, expression levels of IL-4 (Th2 cytokine) did not differ significantly among the different groups throughout the study period. Our findings indicate clear differences in peripheral blood cytokine gene expression levels during pregnancy between animals naturally infected with N. caninum and seronegative control animals. To the best of our knowledge, this is the first study to examine the gene expression of Th1, Th2 and regulatory cytokines in the peripheral blood of pregnant cows naturally infected with N. caninum.

Presence of Toxoplasma gondii and Neospora caninum DNA in the brain of wild birds.

Toxoplasma gondii infections are prevalent in many avian species and can cause mortality in some bird hosts. Although T. gondii has been isolated from various species of birds, the role of many different species of wild birds in the epidemiology of T. gondii remains unknown. Neospora caninum, a closely related parasite to T. gondii, has been recently confirmed to infect domestic chickens and wild birds such as house sparrows (Passer domesticus). The present study reports the presence of T. gondii and N. caninum DNA by PCR in brain tissues of 14 species of wild birds from Spain. From a total of 200 samples analyzed, 12 samples (6%) were positive for T. gondii [5 Eurasian jays (Garrulus glandarius), 5 magpies (Pica pica), 1 black kite (Milvus migrans) and 1 Griffon vulture (Gyps fulvus)], while 3 samples (1.5%) were positive for N. caninum [2 magpies and 1 common buzzard (Buteo buteo)]. This is the first report of detection of T. gondii in magpies, griffon vulture and black kite and of N. caninum in common buzzard and magpies, extending the list of natural intermediate hosts for T. gondii and N. caninum infections to these species.

High seroprevalence of Toxoplasma gondii and Neospora caninum in the Common raven (Corvus corax) in the Northeast of Spain.

In recent years, multiple cases of aggressive behavior of Common ravens (Corvus corax) have been reported by farmers in Catalonia (NE Spain), including attacking of newborn animals and consumption of dead foetuses. In the present study, seroprevalence of Toxoplasma gondii and Neospora caninum was determined from 113 legally trapped and released Common ravens. T. gondii antibodies were found in 91 (80.5%; CI 95%:72-87) of 113 sera tested by the modified agglutination test. Antibodies to N. caninum were found in 24 (35.8%; IC 95%: 24.5-48.5) of 67 Common ravens tested by an indirect fluorescence antibody test with titers ranging from 1:50 (n=18) to ≥1:100 (n=6). To the author's knowledge, this is the first report of the presence of T. gondii and N. caninum antibodies in C. corax. The seroprevalence detected is one of the highest reported worldwide in wild birds, suggesting an important role for this species in the epidemiology of both parasites.

Integument Mycobiota of Wild European Hedgehogs (Erinaceus europaeus) from Catalonia, Spain.

There are some reports about the risk of manipulating wild hedgehogs since they can be reservoirs of potential zoonotic agents like dermatophytes. The aim of this study was to describe the integument mycobiota, with special attention to dermatophytes of wild European hedgehogs. Samples from spines and fur were cultured separately in Sabouraud dextrose agar (SDA) with antibiotic and dermatophyte test medium (DTM) plates. Nineteen different fungal genera were isolated from 91 cultures of 102 hedgehogs. The most prevalent genera were Cladosporium (79.1%), Penicillium (74.7%), Alternaria (64.8%), and Rhizopus (63.7%). A lower prevalence of Aspergillus (P = 0,035; χ (2) = 8,633) and Arthrinium (P = 0,043; χ (2) = 8,173) was isolated during the spring time and higher frequencies of Fusarium (P = 0,015; χ (2) = 10,533) during the autumn. The prevalence of Acremonium was significantly higher in young animals (70%, 26/37) than in adults (30%, 11/37) (P = 0,019; χ (2) = 5,915). Moreover, the majority of the saprophytic species that grew at the SDA culture were also detected at the DTM. Finally, no cases of ringworm were diagnosed and no dermatophytes spp. were isolated. Concluding, this study provides the first description of fungal mycobiota of the integument of wild European hedgehogs in Spain, showing a large number of saprophytic species and the absence of dermatophytes.

Cytokine gene expression at the materno-foetal interface after experimental Neospora caninum infection of heifers at 110 days of gestation.

Neospora caninum is a major cause of abortion in cattle, but the reasons why only some animals abort remain unclear. The immunological control of the parasite in the placenta or by the foetus could be the key to determining the mechanism of abortion and/or transplacental transmission to the foetus. In this study, cytokine gene expression, analysed by real-time RT-PCR, at the maternal (caruncle) and foetal placenta (cotyledon) of heifers infected at 110 days of gestation by intravenous inoculation of N. caninum tachyzoites was compared with the responses in uninfected heifers. Animals were euthanized 3 weeks after infection. Upregulated Th1, Th2 and T-regulatory (Treg) cytokine gene expression was observed in both the maternal and the foetal placenta in the infected group. In the caruncle of infected animals, the main changes included upregulation of IFN-γ, IL-12p40, IL-6 and IL-10. In the cotyledon, the main changes included upregulation of IFN-γ and downregulation of TGF-ß, being the later the only cytokine downregulated in the infected group. The observed cytokine expression pattern was associated with alive but transplacentally infected foetuses, suggesting that such cytokine pattern is beneficial to foetal survival, but could have a role in the transplacental transmission of the parasite.

Human papillomavirus HPV-16, 18, 52 and 58 integration in cervical cells of HIV-1-infected women.

BACKGROUND: Genomic integration of high-risk human papillomavirus into the cellular genome is considered an important event in the pathogenesis of cervical cancer related to the progression from premalignant cervical lesions to invasive cervical carcinoma. OBJECTIVE: This cross-sectional study was aimed to characterize the viral integration of HPV-16, HPV-18, HPV-52 and HPV-58 in cervical cells. STUDY DESIGN: HPV genotypes were determined by PCR and HPV integration by multiplex PCR in HIV-1-infected women without a background of HPV-related pathology. RESULTS: This study included 251 cervical cells samples of consecutive HIV-positive women who were visited between 1999 and 2003. The overall prevalence of HPV infection was 53% (133/251, 95%CI: 47-59%). The most prevalent genotypes were HPV-16 (27%), HPV-33 (15%), HPV-52 (8%) and HPV-58 (8%). The prevalence of abnormal cervical cytology was 33% (83/251, 95%CI: 27-39%). The overall prevalence of HPV integration was 11% (27/251, 95%CI: 7-15%), and the prevalence of HPV-16 integration was 33% (22/67, 95%CI: 22-45%), HPV-18 integration was 30% (3/10, 95%CI: 7-65%) and HPV-52 integration was 10% (2/19, 95%CI: 1-32%). No HPV-58 integration was detected. The percentage of HPV-16 and HPV-18 integration increased with the severity of the cervical lesions, HPV-16 integration was almost 70% and HPV-18 integration was 50% in high-grade squamous intraepithelial lesions. Integration was the most important risk factor associated with cervical dysplasia (OR=30.6, 95%CI: 3.5-270.6). CONCLUSION: HPV integration might represent a good biomarker of the evolution from HPV infection to cervical cancer. Further prospective studies are required to validate our findings.