Post-COVID-19 tomographic abnormalities.

BACKGROUND: The prevalence of persistent respiratory symptoms tends to be low in patients with a longer recovery time after COVID-19. However, some patients may present persistent pulmonary abnormalities.OBJECTIVE: To evaluate the prevalence of tomographic abnormalities 90 days after symptom onset in patients with COVID-19 and compare two chest high-resolution computed tomography (HRCT) analysis techniques.METHODS: A multicentre study of patients hospitalised with COVID-19 having oxygen saturation <93% on room air at hospital admission were evaluated using pulmonary function and HRCT scans 90 days after symptom onset. The images were evaluated by two thoracic radiologists, and were assessed using software that automatically quantified the extent of pulmonary abnormalities.RESULTS: Of the 91 patients included, 81% had at least one pulmonary lobe with abnormalities 90 days after discharge (84% were identified using the automated algorithm). Ground-glass opacities (76%) and parenchymal bands (65%) were the predominant abnormalities. Both chest HRCT technical assessments presented high sensitivity (95.9%) and positive predictive value (92%), with a statistically significant correlation at baseline (R = 0.80) and after 90 days (R = 0.36).CONCLUSION: The prevalence of pulmonary abnormalities on chest HRCT 90 days after symptom onset due to COVID-19 was high; both technical assessments can be used to analyse the images.

Calcicole behaviour of Callisthene fasciculata Mart., an Al-accumulating species from the Brazilian Cerrado.

Most aluminium (Al)-accumulating species are found on soils with high Al saturation and low Ca availability (Ca poor). Callisthene fasciculata Mart. (Vochysiaceae), however, is an Al-accumulating tree restricted to Ca-rich soils with low Al saturation in the Brazilian Cerrado savanna. Here we tested its calcicole behaviour, and the possible role of organic acids in detoxification of Al during the early stages of plant development. We assessed growth, dry mass, nutrients, Al and organic acids in seedlings grown for 50 days on two contrasting Cerrado soils; one with high Ca concentrations and low Al saturation and the other with low Ca availability and high Al saturation. Relative to plants on Ca-rich soil, plants on Ca-poor soil had necrotic spots and bronzing of leaves. Roots and shoots contained reduced concentrations of P and Cu, but higher concentrations of Fe, Al and citrate. Despite lower concentrations in the soil, Ca and Mg increased in shoots. Shoot concentrations of oxalate were also higher. We confirmed C. fasciculata as an Al-accumulating species with calcicole behaviour. The increased concentrations of organic acids in plants with higher Al accumulation suggest that high availability of soluble Al does not prevent occurrence of this species on soils with high Al saturation. Instead, the absence of C. fasciculata from Ca-poor soils is probably due to imbalances in tissue Fe, Cu and Zn imposed by this soil type.

Dispersal mode constrains allocation of carbon and mineral nutrients in seeds of forest and savanna trees.

Species vary in seed size and content of stored reserves, which can be related to dispersal strategies and type of habitat in which they are found. We compare seed carbon and nutrient reserves of anemochorous and zoochorous trees from the Cerrado of central Brazil. We measured seed dry mass, lipids, non-structural carbohydrates (starch and total soluble sugars), carbon and mineral nutrients in ten forest and 13 savanna species, each classified as having wind- or animal-dispersed seeds. We used phylogenetically independent contrasts to test for correlations among these traits. Seeds of anemochorous species were lighter, with higher concentrations of C, N, P, Ca and Mg. Lipids were the dominant carbon reserve for most anemochorous species, underpinning the importance of allocation to compact carbon reserves. Starch, lipids or soluble sugars were the major carbon reserve in zoochorous seeds. Savanna and forest species did not differ in seed mass or in total carbon reserves. However, seeds of forest species had higher concentrations of starch than seeds of savanna species. Lipid and starch negatively correlated across species, suggesting a trade-off between starch and lipids as major seed carbon reserves. Calcium was positively correlated with Mn and B, while Mg was positively correlated with C, N, P, K, S, Zn and B. Potassium, S and Cl were positively correlated, while P was positively correlated with Mg and Zn. Dispersal mode rather than vegetation type constrained seed mass and seed storage allocation patterns in forest and savanna trees. We provide evidence that similar mechanisms are involved in seed storage of carbon and mineral nutrients across species.

Post-fire reproduction of herbs at a savanna-gallery forest boundary in Distrito Federal, Brazil / Reprodução pós-fogo de espécies do estrato herbáceo em área de transição Cerrado-Mata de Galeria no Distrito Federal, Brasil

In Cerrado, studies of post-fire vegetation recovery show that some herbaceous species are able to flower shortly after fires. However, these were mainly short-term studies that focused on grasslands and savannas. Little is known about the effects of fire on ground layer of forests that border the savannas in Central Brazil. Thus, an accidental burning gave us the opportunity to describe the reproductive activity of the ground layer vegetation after a fire event along a savanna-forest boundary at the IBGE Ecological Reserve, Brasília, Brazil. During the 16-month of the inventory, we registered 170 herbaceous species flowering or fruiting, of which 52 species (31%) may have been influenced by fire that changed their times of reproduction. In the savanna plots reproduction peaked at the end of the rainy season. Of the total number of reproducing species, 90 species occurred only in the savanna and four in the forest. Five herbs were recorded in the forest, savanna and border environments. Late dry season fire probably lead the majority of herbaceous species to have their reproduction spread throughout the study time.(AU)

No Cerrado, estudos de recuperação da vegetação pós-fogo mostram que algumas espécies herbáceas são capazes de florescer logo após queimadas. No entanto, esses foram principalmente estudos de curto prazo, que se concentraram em áreas campestres e savânicas. Pouco se sabe sobre os efeitos do fogo no sub-bosque de matas que margeiam as savanas do Brasil Central. Assim, uma queima acidental nos deu a oportunidade de descrever a atividade reprodutiva pós-fogo do estrato herbáceo ao longo de ecótono cerrado-mata na Reserva Ecológica do IBGE, Brasília, Brasil. Durante os 16 meses do inventário, foram registrados 170 espécies herbáceas com flores ou frutos, das quais 52 espécies (31%) podem ter sido influenciadas pelo fogo, que alterou suas épocas reprodutivas. Nas parcelas de cerrado, a reprodução teve seu pico ao final da estação chuvosa. Do número total de espécies, 90 ocorreram exclusivamente no cerrado e quatro na floresta. Cinco espécies herbáceas foram registradas na floresta, cerrado e borda. O fogo ao final da estação seca pode ter levado a maioria das plantas a se reproduzir através do período de estudo.(AU)

Post-fire reproduction of herbs at a savanna-gallery forest boundary in Distrito Federal, Brazil / Reprodução pós-fogo de espécies do estrato herbáceo em área de transição Cerrado-Mata de Galeria no Distrito Federal, Brasil

Abstract In Cerrado, studies of post-fire vegetation recovery show that some herbaceous species are able to flower shortly after fires. However, these were mainly short-term studies that focused on grasslands and savannas. Little is known about the effects of fire on ground layer of forests that border the savannas in Central Brazil. Thus, an accidental burning gave us the opportunity to describe the reproductive activity of the ground layer vegetation after a fire event along a savanna-forest boundary at the IBGE Ecological Reserve, Brasília, Brazil. During the 16-month of the inventory, we registered 170 herbaceous species flowering or fruiting, of which 52 species (31%) may have been influenced by fire that changed their times of reproduction. In the savanna plots reproduction peaked at the end of the rainy season. Of the total number of reproducing species, 90 species occurred only in the savanna and four in the forest. Five herbs were recorded in the forest, savanna and border environments. Late dry season fire probably lead the majority of herbaceous species to have their reproduction spread throughout the study time.

Resumo No Cerrado, estudos de recuperação da vegetação pós-fogo mostram que algumas espécies herbáceas são capazes de florescer logo após queimadas. No entanto, esses foram principalmente estudos de curto prazo, que se concentraram em áreas campestres e savânicas. Pouco se sabe sobre os efeitos do fogo no sub-bosque de matas que margeiam as savanas do Brasil Central. Assim, uma queima acidental nos deu a oportunidade de descrever a atividade reprodutiva pós-fogo do estrato herbáceo ao longo de ecótono cerrado-mata na Reserva Ecológica do IBGE, Brasília, Brasil. Durante os 16 meses do inventário, foram registrados 170 espécies herbáceas com flores ou frutos, das quais 52 espécies (31%) podem ter sido influenciadas pelo fogo, que alterou suas épocas reprodutivas. Nas parcelas de cerrado, a reprodução teve seu pico ao final da estação chuvosa. Do número total de espécies, 90 ocorreram exclusivamente no cerrado e quatro na floresta. Cinco espécies herbáceas foram registradas na floresta, cerrado e borda. O fogo ao final da estação seca pode ter levado a maioria das plantas a se reproduzir através do período de estudo.

Distribution and genetic diversity of the human polyomaviruses JC and BK in surface water and sewage treatment plant during 2009 in Porto Alegre, Southern Brazil / Distribuição e diversidade genética dos poliomavírus humanos JC e BK em águas superficiais e de estação de tratamento de esgoto durante 2009 em Porto Alegre, sul do Brasil

Abstract The human polyomaviruses JC and BK (JCPyV and BKPyV) are ubiquitous, species-specific viruses that belong to the family Polyomaviridae. These viruses are known to be excreted in human urine, and they are potential indicators of human wastewater contamination. In order to assess the distribution of both JCPyV and BKPyV in urban water samples collected from a sewage treatment plant (STP) and from a canalized water stream of Porto Alegre, Brazil, two nested-PCR assays were optimized and applied to the samples collected. The amplicons obtained were submitted to sequencing, and the sequences were analyzed with sequences of human polyomaviruses previously deposited in GenBank. Twelve out of 30 water samples (40%) were JCPyV positive, whereas six samples (20%) were BKPyV positive. The sequencing results confirmed the presence of JCPyV subtypes 1 and 3, whereas only BKPyV Ia and Ib were found. This study shows for the first time the presence of human polyomaviruses in surface water and in samples collected in a sewage treatment plant in southern Brazil.

Resumo Os poliomavírus humanos JC e BK (JCPyV e BKPyV) são virus ubíquos, espécie-específicos, pertencentes à família Polyomaviridae. Estes vírus são conhecidos por serem excretados pela urina humana, sendo considerados potenciais indicadores de contaminação por águas residuais urbanas. Buscando acessar a distribuição de JCPyV e BKPyV em amostras de águas coletadas de uma estação de tratamento de esgoto e de um arroio canalizado de Porto Alegre, Brasil, duas técnicas de nested-PCR foram otimizadas e aplicadas às amostras coletadas. Os amplificados obtidos foram submetidos ao sequenciamento e suas sequências analisadas com base em sequências de poliomavírus humanos previamente depositadas no GenBank. Doze de 30 amostras de água (40%) foram positivas para JCPyV, enquanto 6 amostras (20%) foram positivas para BKPyV. Os resultados do sequenciamento confirmaram a presença dos subtipos 1 e 3 de JCPyV, enquanto apenas os BKPyV Ia e Ib foram encontrados. Este estudo demonstra pela primeira vez a presença de poliomavírus humanos em águas superficiais e em amostras coletadas em uma estação de tratamento de esgoto na região sul do Brasil.

Post-fire reproduction of herbs at a savanna-gallery forest boundary in Distrito Federal, Brazil.

In Cerrado, studies of post-fire vegetation recovery show that some herbaceous species are able to flower shortly after fires. However, these were mainly short-term studies that focused on grasslands and savannas. Little is known about the effects of fire on ground layer of forests that border the savannas in Central Brazil. Thus, an accidental burning gave us the opportunity to describe the reproductive activity of the ground layer vegetation after a fire event along a savanna-forest boundary at the IBGE Ecological Reserve, Brasília, Brazil. During the 16-month of the inventory, we registered 170 herbaceous species flowering or fruiting, of which 52 species (31%) may have been influenced by fire that changed their times of reproduction. In the savanna plots reproduction peaked at the end of the rainy season. Of the total number of reproducing species, 90 species occurred only in the savanna and four in the forest. Five herbs were recorded in the forest, savanna and border environments. Late dry season fire probably lead the majority of herbaceous species to have their reproduction spread throughout the study time.

Distribution and genetic diversity of the human polyomaviruses JC and BK in surface water and sewage treatment plant during 2009 in Porto Alegre, Southern Brazil.

The human polyomaviruses JC and BK (JCPyV and BKPyV) are ubiquitous, species-specific viruses that belong to the family Polyomaviridae. These viruses are known to be excreted in human urine, and they are potential indicators of human wastewater contamination. In order to assess the distribution of both JCPyV and BKPyV in urban water samples collected from a sewage treatment plant (STP) and from a canalized water stream of Porto Alegre, Brazil, two nested-PCR assays were optimized and applied to the samples collected. The amplicons obtained were submitted to sequencing, and the sequences were analyzed with sequences of human polyomaviruses previously deposited in GenBank. Twelve out of 30 water samples (40%) were JCPyV positive, whereas six samples (20%) were BKPyV positive. The sequencing results confirmed the presence of JCPyV subtypes 1 and 3, whereas only BKPyV Ia and Ib were found. This study shows for the first time the presence of human polyomaviruses in surface water and in samples collected in a sewage treatment plant in southern Brazil.

Genetic divergence among invasive and native populations of the yellow peacock cichlid Cichla kelberi.

This study used the hypervariable domain of the mitochondrial DNA (mtDNA) control region (CR) to assess the genetic divergence among native and invasive populations of Cichla kelberi, which is considered the first peacock cichlid introduced and established throughout Brazil and is among the most invasive populations of this genus worldwide. The maximum likelihood tree based on 53 CR sequences with strong bootstrap support revealed that C. kelberi forms a monophyletic clade, confirming that all 30 C. kelberi studied belong to this morphotype. Additionally, the haplotype analysis of the C. kelberi sequences from 11 sampling sites revealed that invasive populations are much less diverse than native ones and largely dominated by a single haplotype that prevailed in reservoirs at the Paraíba do Sul River basin. Two haplotypes were recorded exclusively in an invasive population at Porto Rico, southern Brazil, and one private haplotype was detected in two reservoirs from Paraíba do Sul (Pereira Passos and Paracambi), suggesting more than one introduction event and that native populations should be better evaluated to encompass the entire genetic diversity of native C. kelberi. The possible route and pathways of C. kelberi introduction are also briefly discussed.

A Spontaneous Deletion of the US1.67/US2 Genomic Region on the Bovine Herpesvirus 1 Strain Cooper.

Bovine herpesvirus 1 (BoHV-1) is an alphaherpesvirus with a genome of 135 kb. Some BoHV-1 genes are nonessential and may be deleted from the viral genome. Here, a spontaneous gene deletion was identified in the BoHV-1 strain Cooper. Genes of the US1.67/US2 region were absent, as determined by next-generation sequencing.

Responses of fish assemblages of sandy beaches to different anthropogenic and hydrodynamic influences.

This study tested whether some attributes of the diversity, communities and populations of surf-zone fish assemblages varied with different hydrodynamic and anthropogenic influences at four Guanabara Bay sandy beaches: Dentro (sheltered with limited human access), Fora (exposed with limited human access), Urca (sheltered with unlimited human access) and Vermelha (exposed with unlimited human access), between autumn 2011 and summer 2012. Twenty-nine species and 1613 individuals were recorded from 76 trawls. The 10 most abundant species accounted for 94·5% of the total number, but only four species (Diplodus argenteus, Harengula clupeola, Sardinella brasiliensis and Sphoeroides greeleyi) were recorded at all four beaches, revealing a high level of species substitution. Fish assemblages differed not only for diversity attributes, but also at community and population levels, with lower values of the Shannon-Wiener index, richness and total fish abundance and biomass at Vermelha beach, and higher densities of Trachinotus carolinus, Atherinella brasiliensis and S. greeleyi related to beaches with high anthropogenic influence. The findings reveal that fish assemblages of Dentro, Fora, Urca and Vermelha beaches differed not only in response to hydrodynamic influences, but also due to the effects of different degrees of human interference (i.e. presence of solid residues, population density and fishing impacts), emphasizing the importance of the sheltered and less anthropogenically affected beaches, as spawning, nursery and growth areas.

Complete Genome Sequence of Porcine Parvovirus 2 Recovered from Swine Sera.

A complete genomic sequence of porcine parvovirus 2 (PPV-2) was detected by viral metagenome analysis on swine sera. A phylogenetic analysis of this genome reveals that it is highly similar to previously reported North American PPV-2 genomes. The complete PPV-2 sequence is 5,426 nucleotides long.

Swine Influenza Virus and Association with the Porcine Respiratory Disease Complex in Pig Farms in Southern Brazil.

Despite the putative endemic status of swine influenza A virus (swIAV) infections, data on the occurrence of swine influenza outbreaks are scarce in Brazil. The aim of this study was to detect and subtype swIAVs from six outbreaks of porcine respiratory disease complex (PRDC) in southern Brazil. Nasal swabs were collected from 66 piglets with signs of respiratory disease in six herds. Lung tissue samples were collected from six necropsied animals. Virus detection was performed by PCR screening and confirmed by virus isolation and hemagglutination (HA). Influenza A subtyping was performed by a real-time reverse transcriptase PCR (rRT-PCR) to detect the A(H1N1)pdm09; other swIAV subtypes were determined by multiplex RT-PCR. In lung tissues, the major bacterial and viral pathogens associated with PRDC (Pasteurella multocida, Mycoplasma hyopneumoniae, Actinobacillus pleuropneumoniae, Haemophilus parasuis and PCV2) were investigated. In some affected pigs, clinico-pathological evaluations were conducted. Influenza A was detected by screening PCR in 46 of 66 swab samples and from five of six lungs. Virus was recovered from pigs of all six herds. Subtype A(H1N1)pdm09 was detected in four of six herds and H1N2 in the other two herds. In lung tissues, further agents involved in PRDC were detected in all cases; Pasteurella multocida was identified in five of six samples and Mycoplasma hyopneumoniae in three of six. Actinobacillus pleuropneumoniae (1/6), Haemophilus parasuis (1/6) and PCV2 (1/6) were also detected. These findings indicate that subtypes A(H1N1)pdm09 and H1N2 were present in pigs in southern Brazil and were associated with PRDC outbreaks.

Coccinellidae Parasitoids in Brazil: Neglected Species in a Mega-Diverse Country.

Current knowledge on coccinellids is primarily focused on their role as natural enemies of soft-bodied insects. However, there is a great diversity of coccinellid parasitoid species that are less studied. Here, we describe new records of coccinellid parasitoids with emphasis on new host-parasitoid interactions in 11 sample sites in Brazil. We collected 122 coccinellid individuals parasitized by six species of parasitoids in the Cerrado and in the Atlantic Rainforest biomes. New records of coccinellid parasitoids and host associations, expansion of habitat ranges and interactions are discussed focusing on the lack of basic information on these interactions in Brazil.

A comparative study of aluminium and nutrient concentrations in mistletoes on aluminium-accumulating and non-accumulating hosts.

Mistletoes offer a unique model to study interactions among Al and nutrients in vascular plants, because they grow and reproduce on hosts with distinct Al uptake strategies. We investigated Al distribution and nutrient relations of mistletoes on Al-accumulating and non-accumulating hosts. We hypothesised that mistletoes would exhibit similar leaf nutrient and Al concentrations as their host plants, but a strong compartmentalisation of Al when growing on Al-accumulators. We measured concentrations of N, P, K, Ca, Mg, Cu, Fe, Mn, Zn in leaves and Al in leaves, seeds and branches of Phthirusa ovata and Psittacanthus robustus infecting Miconia albicans, an Al-accumulator, and Ph. ovata infecting Byrsonima verbascifolia, a non-Al-accumulator. High leaf concentrations of Al in Ph. ovata only occurred while parasitizing the Al-accumulating host; there was no accumulation in branches or seeds. In P. robustus, large concentrations of Al were found in leaves, branches and seeds. Mistletoe seed viability and leaf nutrient concentrations were not affected by Al accumulation. Passive uptake of Al, Ca, Mg, Mn and Cu in mistletoes was evidenced by significant correlations between mistletoes and host leaf concentrations, but not of N, P and K. Al was retranslocated to different plant organs in P. robustus, whereas it was mostly restricted to leaves in Ph. ovata. We suggest that Al might have some specific function in P. robustus, which only parasitizes Al-accumulator hosts, while the host generalist Ph. ovata can be considered a facultative Al-accumulator.

The partitioning of water uptake between growth forms in a Neotropical savanna: do herbs exploit a third water source niche?

In addition to trees and grasses, the savannas of central Brazil are characterised by a diverse herbaceous dicot flora. Here we tested whether the coexistence of a highly diversified assemblage of species resulted in stratification or strong overlap in the use of soil water resources. We measured oxygen and hydrogen isotope ratios of stem water from herbs, grasses and trees growing side by side, as well as the isotopic composition of water in soil profile, groundwater and rainfall, and predawn (Ψ(pd)) and midday (Ψ(md)) leaf water potentials. We used a stable isotope mixing model to estimate vertical partitioning of soil water by the three growth forms. Grasses relied on shallow soil water (5-50 cm) and were strongly anisohydric. Ψ(pd) and Ψ(md) decreased significantly from the wet to the dry season. Trees extracted water from deeper regions of the soil profile (60-120 cm) and were isohydric. Ψ(pd) and Ψ(md) did not change from the wet to the dry season. Herbs overlapped with grasses in patterns of water extraction in the dry season (between 10 and 40 cm), but they took up water at soil depths intermediate (70-100 cm) to those of trees and grasses during the wet season. They showed seasonal changes in Ψ(pd) but not in Ψ(md). We conclude that vertical partitioning of soil water may have contributed to coexistence of these three growth forms and resulted in a more complex pattern of soil water extraction than the two-compartment model of soil water uptake currently used to explain the structure and function of tropical savanna ecosystems.

First description of Adenovirus, Enterovirus, Rotavirus and Torque teno virus in water samples collected from the Arroio Dilúvio, Porto Alegre, Brazil.

Adenovirus (AdV), enterovirus (EV), genogroup A rotaviruses (GARV) and Torque teno virus (TTV) are non-enveloped viral agents excreted in feces and so may contaminate water bodies. In the present study, the molecular detection of these viruses was performed in samples of surface water collected from the Arroio Dilúvio, a waterstream that crosses the city of Porto Alegre, RS, Brazil, receiving great volumes of non-treated sewage from a large urban area. Sampling was performed during 2009, in three different occasions (January, April and September). The highest detection rate was observed for EV (64.28%), followed by TTV (28.57%) and AdV (21.43%). Rotaviruses were not detected. More than on kind of tested virus was detected in five (35. 71%) of 14 samples. January was the month with the highest viral detection rate, being all samples, collected in this month, positive for at least one group of tested virus. The correlation between the detection of these different viral agents and environmental factors is discussed. To the knowledge of the authors, this is the first description of viral genomes in water samples taken from the Arroio Dilúvio, Porto Alegre (Brazil).

Immunoperoxidase inhibition assay for rabies antibody detection.

An immunoperoxidase inhibition assay (IIA) for detection of rabies antibodies in human sera is described. Diluted test sera are added to microplates with paraformaldehyde-fixed, CER cells infected with rabies virus. Antibodies in test sera compete with a rabies polyclonal rabbit antiserum which was added subsequently. Next, an anti-rabbit IgG-peroxidase conjugate is added and the reaction developed by the addition of the substrate 3-amino-9-ethylcarbazole (AEC). The performance of the assay was compared to that of the "simplified fluorescence inhibition microtest" (SFIMT), an established virus neutralization assay, by testing 422 human sera. The IIA displayed 97.6% sensitivity, 98% specificity and 97.6% accuracy (Kappa correlation coefficient=0.9). The IIA results can be read by standard light microscopy, where the clearly identifiable specific staining is visible in antibody-negative sera, in contrast to the absence of staining in antibody-positive samples. The assay does not require monoclonal antibodies or production of large amounts of virus; furthermore, protein purification steps or specialized equipment are not necessary for its performance. The IIA was shown to be suitable for detection of rabies antibodies in human sera, with sensitivity, specificity and accuracy comparable to that of a neutralization-based assay. This assay may be advantageous over other similar methods designed to detect rabies-specific binding antibodies, in that it can be easily introduced into laboratories, provided basic cell culture facilities are available.

Detection of bovine herpesvirus 1 and 5 in semen from Brazilian bulls.

Bovine herpesvirus 1 (BoHV-1) and 5 (BoHV-5) are important pathogens of the respiratory and genital tract of cattle and may also affect the central nervous system and cause meningoencephalitis. Both virus types are estimated to be widely distributed in Southern Brazil. In the present study, BoHV-1 and/or BoHV-5 DNA were detected in bovine semen samples from two states of Brazil by two species-specific nested polymerase chain reactions (nPCRs). These nPCRs were used to assay 53 samples of fresh semen and 23 samples of frozen semen from breeding bulls. Viral DNA was detected in all 76 semen samples: all were positive for BoHV-5, whereas 34 of these were positive for BoHV-1 as well. Moreover, in five fresh and in 13 frozen semen samples-of a total number of 40 samples suitable for virus isolation-infectious BoHV-1 and/or BoHV-5 virus were detected. In conclusion, that both BoHV-1 and BoHV-5 were detected in bovine semen in Brazil highlighted the importance of examining bull semen in search for both agents to reduce the risk of transmitting these viruses.

Efficacy of an inactivated, recombinant bovine herpesvirus type 5 (BoHV-5) vaccine.

Bovine herpesvirus type 5 (BoHV-5) is the causative agent of bovine herpetic encephalitis. In countries where BoHV-5 is prevalent, attempts to vaccinate cattle to prevent clinical signs from BoHV-5-induced disease have relied essentially on vaccination with BoHV-1 vaccines. However, such practice has been shown not to confer full protection to BoHV-5 challenge. In the present study, an inactivated, oil adjuvanted vaccine prepared with a recombinant BoHV-5 from which the genes coding for glycoprotein I (gI), glycoprotein E (gE) and membrane protein US9 were deleted (BoHV-5 gI/gE/US9(-)), was evaluated in cattle in a vaccination/challenge experiment. The vaccine was prepared from a virus suspension containing a pre-inactivation antigenic mass equivalent to 10(7.69) TCID(50)/dose. Three mL of the inactivated vaccine were administered subcutaneously to eight calves serologically negative for BoHV-5 (vaccinated group). Four other calves were mock-vaccinated with an equivalent preparation without viral antigens (control group). Both groups were boostered 28 days later. Neither clinical signs of disease nor adverse effects were observed during or after vaccination. A specific serological response, revealed by the development of neutralizing antibodies, was detected in all vaccinated animals after the first dose of vaccine, whereas control animals remained seronegative. Calves were subsequently challenged on day 77 post-vaccination (pv) with 10(9.25) TCID(50) of the wild-type BoHV-5 (parental strain EVI 88/95). After challenge, vaccinated cattle displayed mild signs of respiratory disease, whereas the control group developed respiratory disease and severe encephalitis, which led to culling of 2/4 calves. Searches for viral DNA in the central nervous system (CNS) of vaccinated calves indicated that wild-type BoHV-5 did not replicate, whereas in CNS tissues of calves on the control group, viral DNA was widely distributed. BoHV-5 shedding in nasal secretions was significantly lower in vaccinated calves than in the control group on days 2, 3, 4 and 6 post-challenge (pc). In addition, the duration of virus shedding was significantly shorter in the vaccinated (7 days) than in controls (12 days). Attempts to reactivate latent infection by administration of dexamethasone at 147 days pv led to recrudescence of mild signs of respiratory disease in both vaccinated and control groups. Infectious virus shedding in nasal secretions was detected at reactivation and was significantly lower in vaccinated cattle than in controls on days 11-13 post-reactivation (pr). It is concluded that the inactivated vaccine prepared with the BoHV-5 gI/gE/US9(-) recombinant was capable of conferring protection to encephalitis when vaccinated cattle were challenged with a large infectious dose of the parental wild type BoHV-5. However, it did not avoid the establishment of latency nor impeded dexamethasone-induced reactivation of the virus, despite a significant reduction in virus shedding after challenge and at reactivation on vaccinated calves.