Factors Influencing Milk Quality and Subclinical Mastitis in Dairy Herds Housed in Compost-Bedded Pack Barn System.

The compost-bedded pack barn (CBPB) system has been increasingly adopted by dairy farms due to its ability to enhance animal comfort and milk production. This study evaluated the associations among bedding characteristics, milk quality and composition, and subclinical mastitis (SCM) occurrence in dairy herds housed in CBPB systems. Over a period of six months, data related to milk quality and udder health and bedding sampling were collected from eight dairy farms. Monthly measurements of the bedding temperature and wind speed inside the CBPB were taken, while temperature and relative humidity data inside the CBPB were recorded using a datalogger. Bedding samples were subjected to analysis of moisture, pH, microbiological count, and carbon/nitrogen ratio. Data on milk composition (fat, protein, milk urea nitrogen, and total solids) and quality (somatic cell count and standard plate count) of bulk tank milk were obtained from DHIA results. Canonical correlation analyses were used to evaluate the association between the analyzed group variables, and linear regression models were used to identify associations between bedding characteristics and SCM occurrence in the studied herds. The bedding characteristics that most influenced milk composition and quality were moisture, temperature at 30 cm depth (T30), and bedding pH. Environmental variables played an important role in bedding composting, as they were closely related to the surface temperature and pH. Overall, 62.71% of the variation in milk quality and composition could be explained by the bedding variables, and 77.50% of the variation in the bedding variables was associated with environmental variables. Median SCM prevalence and incidence were 28.6 and 13.8%, respectively. An increase of 1 °C for T30 resulted in a 0.6% reduction in the prevalence of SCM. Additionally, the bedding surface temperature at 22.3 °C resulted in the highest incidence of SCM (~18.1%). Our results demonstrate the importance of controlling microclimatic conditions in the CBPB to optimize the bedding composting process and milk quality.

Spatial variability of chemical indicators of Amazon agricultural soils through geomultivariate statistics, Brazil.

This work focuses on evaluating the spatial variability of chemical attributes of soils under different agricultural use and native forest, indicating which are the possible indicator attributes of changes in environmental, through the use and management of the soil. The study was carried out in the southern region of the Amazonas state, in an Argissolo Vermelho-Amarelo (Ultisol). Sampling grids were established measuring: 90 m × 70 m with regular soil collection spacing of 10 m for the guarana and forest areas; 90 m × 56 m spaced at 10 m × 8 m for annatto area; and 54 m × 42 m with spacing between points of 6 m for the cupuaçu area, totaling 80 sampling points in each area, with soil samples collected at depths of 0.0-0.05; 0.05-0.10 m and 0.10-0.20 m. The following attributes were determined: pH, Al3+, K+, Ca2+, Mg2+, P, H + Al, CEC, V% and m%. Descriptive, geostatistical and multivariate statistical analyzes were performed. The results show that it is possible to state that the descriptive, geostatistical and multivariate statistical techniques were able to identify the difference between the spatial variability of the attributes according to each specific use of individual soils. The multivariate analysis made it possible to select the attributes that most contribute to the variability of these soils, and with that, it was found that the forest showed less spatial variability in the surface layer, with higher reach values by scaled semivariograms.

Coral microbiome manipulation elicits metabolic and genetic restructuring to mitigate heat stress and evade mortality.

Beneficial microorganisms for corals (BMCs) ameliorate environmental stress, but whether they can prevent mortality and the underlying host response mechanisms remains elusive. Here, we conducted omics analyses on the coral Mussismilia hispida exposed to bleaching conditions in a long-term mesocosm experiment and inoculated with a selected BMC consortium or a saline solution placebo. All corals were affected by heat stress, but the observed "post-heat stress disorder" was mitigated by BMCs, signified by patterns of dimethylsulfoniopropionate degradation, lipid maintenance, and coral host transcriptional reprogramming of cellular restructuration, repair, stress protection, and immune genes, concomitant with a 40% survival rate increase and stable photosynthetic performance by the endosymbiotic algae. This study provides insights into the responses that underlie probiotic host manipulation. We demonstrate that BMCs trigger a dynamic microbiome restructuring process that instigates genetic and metabolic alterations in the coral host that eventually mitigate coral bleaching and mortality.

Pseudorandom full-field electroretinograms reflect different light adaptation mechanisms.

PURPOSE: To investigate the magnitude and time course of pseudorandom ffERG during light adaptation. METHODS: Ten healthy subjects (26 ± 10.1 years) underwent 20 min of dark adaptation, and then the ffERG was evoked by pseudorandom flash sequences (4 ms per flash, 3 cd.s/m2) driven by m-sequences (210-1 stimulus steps) using Veris Science software and a Ganzfeld dome over a constant field of light adaptation (30 cd/m2). The base period of the m-sequence was 50 ms. Each stimulation sequence lasting 40 s was repeated at 0, 5, 10, 15 and 20 min of light adaptation. Relative amplitude and latency (corrected by values found at 0 min) of the three components (N1, P1, and N2) of first-order (K1) and first slice of the second-order (K2.1) kernel at 5 time points were evaluated. An exponential model was fitted to the mean amplitude and latency data as a function of the light adaptation duration to estimate the time course (τ) of the light adaptation for each component. Repeated one-way ANOVA followed by Tukey post-test was applied to the amplitude and latency data, considering significant values of p < 0.05. RESULTS: Regarding the K1 ffERG, N1 K1, P1 K1, and N2 K1 presented an amplitude increase as a function of the light adaptation (N1 K1 τ value = 2.66 min ± 4.2; P1 K1 τ value = 2.69 min ± 2.10; and N2 K1 τ value = 3.49 min ± 2.96). P1 K1 and N2 K1 implicit time changed as a function of the light adaptation duration (P1 K1 τ value = 3.61 min ± 5.2; N2 K1 τ value = 3.25 min ± 4.8). N1 K1 had small implicit time changes during the light adaptation. All the K2,1 components also had nonsignificant changes in amplitude and implicit time during the light adaptation. CONCLUSIONS: Pseudorandom ffERGs showed different mechanisms of adaptation to retinal light. Our results suggest that K1 ffERG is generated by retinal mechanisms with intermediate- to long-term light adaptation, while K2.1 ffERG is generated by retinal mechanism with fast light adaptation course.

Delivering Beneficial Microorganisms for Corals: Rotifers as Carriers of Probiotic Bacteria.

The use of Beneficial Microorganisms for Corals (BMCs) to increase the resistance of corals to environmental stress has proven to be effective in laboratory trials. Because direct inoculation of BMCs in larger tanks or in the field can be challenging, a delivery mechanism is needed for efficient transmission of the BMC consortium. Packaged delivery mechanisms have been successfully used to transmit probiotics to other organisms, including humans, lobsters, and fish. Here, we tested a method for utilizing rotifers of the species Brachionus plicatilis for delivery of BMCs to corals of the species Pocillopora damicornis. Epifluorescence microscopy combined with a live/dead cell staining assay was used to evaluate the viability of the BMCs and monitor their in vivo uptake by the rotifers. The rotifers efficiently ingested BMCs, which accumulated in the digestive system and on the body surface after 10 min of interaction. Scanning electron microscopy confirmed the adherence of BMCs to the rotifer surfaces. BMC-enriched rotifers were actively ingested by P. damicornis corals, indicating that this is a promising technique for administering coral probiotics in situ. Studies to track the delivery of probiotics through carriers such as B. plicatilis, and the provision or establishment of beneficial traits in corals are the next proof-of-concept research priorities.

Prospecting Microbial Strains for Bioremediation and Probiotics Development for Metaorganism Research and Preservation.

Pollution affects all biomes. Marine environments have been particularly impacted, especially coral reefs, one of the most sensitive ecosystems on Earth. Globally, 4.5 billion people are economically dependent on the sea, where most of their livelihood is provided by coral reefs. Corals are of great importance and therefore their extinction leads to catastrophic consequences. There are several possible solutions to remediate marine pollutants and local contamination, including bioremediation. Bioremediation is the capacity of organisms to degrade contaminants. The approach presents several advantages, such as sustainability, relatively low cost, and the fact that it can be applied in different ecosystems, causing minimal impacts to the environment. As an extra advantage, the manipulation of endogenous microbiomes, including putative beneficial microorganisms for corals (pBMCs), may have probiotic effects for marine animals. In this context, the use of the two approaches, bioremediation and pBMC inoculation combined, could be promising. This strategy would promote the degradation of specific pollutants that can be harmful to corals and other metaorganisms while also increasing host resistance and resilience to deal with pollution and other threats. This method focuses on the selection of pBMCs to degrade two contaminants: the synthetic estrogen 17a-ethinylestradiol (EE2) and crude oil. Both have been reported to negatively impact marine animals, including corals, and humans. The protocol describes how to isolate and test bacteria capable of degrading the specific contaminants, followed by a description of how to detect some putative beneficial characteristics of these associated microbes to their coral host. The methodologies described here are relatively cheap, easy to perform, and highly adaptable. Almost any kind of soluble target compound can be used instead of EE2 and oil.

Global network of computational biology communities: ISCB's Regional Student Groups breaking barriers.

Regional Student Groups (RSGs) of the International Society for Computational Biology Student Council (ISCB-SC) have been instrumental to connect computational biologists globally and to create more awareness about bioinformatics education. This article highlights the initiatives carried out by the RSGs both nationally and internationally to strengthen the present and future of the bioinformatics community. Moreover, we discuss the future directions the organization will take and the challenges to advance further in the ISCB-SC main mission: "Nurture the new generation of computational biologists".

KSG estimation of reconstruction delay to detect vocal disorders in nonlinear dynamical analysis

Abstract Introduction This research investigates the applicability of a relatively new estimator of mutual information, KSG estimator, to find the reconstruction delay of phase space in dynamical systems. There are evidences that the KSG estimator is more accurate than the naive method commonly used. Methods In this paper we estimated mutual information between the voice signals and their delayed versions, with KSG method. The voice signals were obtained from a disordered voice database. Then, we found the reconstruction delay where mutual information reached its first minimum. We applied the encountered value of reconstruction delay in linear discriminant analysis, in order to discriminate between healthy and pathological voices or to discriminate between pathologies. Discrimination between voice pathologies using nonlinear measurements is still not much explored. Moreover, in this paper we used a single nonlinear measurement: reconstruction delay. Results The results show that the reconstruction delay obtained with KSG method has increased classification rates in most cases, in terms of accuracy, sensitivity and specificity, when compared to the naive estimator usually adopted. Conclusion The KSG estimator is a promising technique to improve the diagnosis of voice related pathologies.

Absorption and translocation of sulfometuron-methyl in sugarcane (Saccharum officinarum L.) at different growth stages.

In Brazil, weed management in sugarcane fields is mainly done with the use of selective herbicide formulations. For many years, diuron+hexazinone was one of the main herbicide mixture formulations used in sugarcane. Later, sulfometuron-methyl was included in the same mixture, which was marketed as a new herbicide formulation for residual in-season weed control in sugarcane. The mixture diuron+hexazinone+sulfometuron-methyl has been widely used in commercial sugarcane fields in Brazil. However, recent field observations have shown that sugarcane plants at different growth stages varied in their phytotoxicity levels after treatment with diuron+hexazinone+sulfometuron-methyl. Greenhouse and laboratory studies were conducted to determine 14Csulfometuron-methyl absorption and translocation, as well as 14C distribution in sugarcane at two growth stages, 2 to 3 leaves and 5 to 6 leaves. 14Csulfometuron-methyl absorption by sugarcane did not differ between the two growth stages. Different patterns of 14C accumulation were observed, which may explain variations in sulfometuron-methyl phytotoxic responses observed in the field.

Genome-wide signatures of complex introgression and adaptive evolution in the big cats.

The great cats of the genus Panthera comprise a recent radiation whose evolutionary history is poorly understood. Their rapid diversification poses challenges to resolving their phylogeny while offering opportunities to investigate the historical dynamics of adaptive divergence. We report the sequence, de novo assembly, and annotation of the jaguar (Panthera onca) genome, a novel genome sequence for the leopard (Panthera pardus), and comparative analyses encompassing all living Panthera species. Demographic reconstructions indicated that all of these species have experienced variable episodes of population decline during the Pleistocene, ultimately leading to small effective sizes in present-day genomes. We observed pervasive genealogical discordance across Panthera genomes, caused by both incomplete lineage sorting and complex patterns of historical interspecific hybridization. We identified multiple signatures of species-specific positive selection, affecting genes involved in craniofacial and limb development, protein metabolism, hypoxia, reproduction, pigmentation, and sensory perception. There was remarkable concordance in pathways enriched in genomic segments implicated in interspecies introgression and in positive selection, suggesting that these processes were connected. We tested this hypothesis by developing exome capture probes targeting ~19,000 Panthera genes and applying them to 30 wild-caught jaguars. We found at least two genes (DOCK3 and COL4A5, both related to optic nerve development) bearing significant signatures of interspecies introgression and within-species positive selection. These findings indicate that post-speciation admixture has contributed genetic material that facilitated the adaptive evolution of big cat lineages.

Corrigendum: Whole genome analysis of a schistosomiasis-transmitting freshwater snail.

This corrects the article DOI: 10.1038/ncomms15451.

Whole genome analysis of a schistosomiasis-transmitting freshwater snail.

Biomphalaria snails are instrumental in transmission of the human blood fluke Schistosoma mansoni. With the World Health Organization's goal to eliminate schistosomiasis as a global health problem by 2025, there is now renewed emphasis on snail control. Here, we characterize the genome of Biomphalaria glabrata, a lophotrochozoan protostome, and provide timely and important information on snail biology. We describe aspects of phero-perception, stress responses, immune function and regulation of gene expression that support the persistence of B. glabrata in the field and may define this species as a suitable snail host for S. mansoni. We identify several potential targets for developing novel control measures aimed at reducing snail-mediated transmission of schistosomiasis.

The Echinococcus canadensis (G7) genome: a key knowledge of parasitic platyhelminth human diseases.

BACKGROUND: The parasite Echinococcus canadensis (G7) (phylum Platyhelminthes, class Cestoda) is one of the causative agents of echinococcosis. Echinococcosis is a worldwide chronic zoonosis affecting humans as well as domestic and wild mammals, which has been reported as a prioritized neglected disease by the World Health Organisation. No genomic data, comparative genomic analyses or efficient therapeutic and diagnostic tools are available for this severe disease. The information presented in this study will help to understand the peculiar biological characters and to design species-specific control tools. RESULTS: We sequenced, assembled and annotated the 115-Mb genome of E. canadensis (G7). Comparative genomic analyses using whole genome data of three Echinococcus species not only confirmed the status of E. canadensis (G7) as a separate species but also demonstrated a high nucleotide sequences divergence in relation to E. granulosus (G1). The E. canadensis (G7) genome contains 11,449 genes with a core set of 881 orthologs shared among five cestode species. Comparative genomics revealed that there are more single nucleotide polymorphisms (SNPs) between E. canadensis (G7) and E. granulosus (G1) than between E. canadensis (G7) and E. multilocularis. This result was unexpected since E. canadensis (G7) and E. granulosus (G1) were considered to belong to the species complex E. granulosus sensu lato. We described SNPs in known drug targets and metabolism genes in the E. canadensis (G7) genome. Regarding gene regulation, we analysed three particular features: CpG island distribution along the three Echinococcus genomes, DNA methylation system and small RNA pathway. The results suggest the occurrence of yet unknown gene regulation mechanisms in Echinococcus. CONCLUSIONS: This is the first work that addresses Echinococcus comparative genomics. The resources presented here will promote the study of mechanisms of parasite development as well as new tools for drug discovery. The availability of a high-quality genome assembly is critical for fully exploring the biology of a pathogenic organism. The E. canadensis (G7) genome presented in this study provides a unique opportunity to address the genetic diversity among the genus Echinococcus and its particular developmental features. At present, there is no unequivocal taxonomic classification of Echinococcus species; however, the genome-wide SNPs analysis performed here revealed the phylogenetic distance among these three Echinococcus species. Additional cestode genomes need to be sequenced to be able to resolve their phylogeny.

Whole genome sequencing of Guzerá cattle reveals genetic variants in candidate genes for production, disease resistance, and heat tolerance.

In bovines, artificial selection has produced a large number of breeds which differ in production, environmental adaptation, and health characteristics. To investigate the genetic basis of these phenotypical differences, several bovine breeds have been sequenced. Millions of new SNVs were described at every new breed sequenced, suggesting that every breed should be sequenced. Guzerat or Guzerá is an indicine breed resistant to drought and parasites that has been the base for some important breeds such as Brahman. Here, we describe the sequence of the Guzerá genome and the in silico functional analyses of intragenic breed-specific variations. Mate-paired libraries were generated using the ABI SOLiD system. Sequences were mapped to the Bos taurus reference genome (UMD 3.1) and 87% of the reference genome was covered at a 26X. Among the variants identified, 2,676,067 SNVs and 463,158 INDELs were homozygous, not found in any database searched, and may represent true differences between Guzerá and B. taurus. Functional analyses investigated with the NGS-SNP package focused on 1069 new, non-synonymous SNVs, splice-site variants (including acceptor and donor sites, and the conserved regions at both intron borders, referred to here as splice regions) and coding INDELs (NS/SS/I). These NS/SS/I map to 935 genes belonging to cell communication, environmental adaptation, signal transduction, sensory, and immune systems pathways. These pathways have been involved in phenotypes related to health, adaptation to the environment and behavior, and particularly, disease resistance and heat tolerance. Indeed, 105 of these genes are known QTLs for milk, meat and carcass, production, reproduction, and health traits. Therefore, in addition to describing new genetic variants, our approach provided groundwork for unraveling key candidate genes and mutations.

Auditory Stimuli Coding by Postsynaptic Potential and Local Field Potential Features.

The relation between physical stimuli and neurophysiological responses, such as action potentials (spikes) and Local Field Potentials (LFP), has recently been experimented in order to explain how neurons encode auditory information. However, none of these experiments presented analyses with postsynaptic potentials (PSPs). In the present study, we have estimated information values between auditory stimuli and amplitudes/latencies of PSPs and LFPs in anesthetized rats in vivo. To obtain these values, a new method of information estimation was used. This method produced more accurate estimates than those obtained by using the traditional binning method; a fact that was corroborated by simulated data. The traditional binning method could not certainly impart such accuracy even when adjusted by quadratic extrapolation. We found that the information obtained from LFP amplitude variation was significantly greater than the information obtained from PSP amplitude variation. This confirms the fact that LFP reflects the action of many PSPs. Results have shown that the auditory cortex codes more information of stimuli frequency with slow oscillations in groups of neurons than it does with slow oscillations in neurons separately.

[Canine visceral leishmaniasis diagnosis by immunohistochemistry and PCR in skin tissues in association with IFAT and ELISA-test]. / Diagnóstico da Leishmaniose Visceral Canina pelas técnicas de imunoistoquímica e PCR em tecidos cutâneos em associação com a RIFI e ELISA-teste.

The purpose of the present study was to evaluate the immunohistochemistry (IMHC) and PCR (Polymerase Chain Reaction) tests for Canine Visceral Leishmaniasis (CVL) diagnosis and compare the results with serological tests such as the indirect fluorescence antibody test (IFAT), ELISA and a parasitological test (microscopic direct examination of the parasite stained with haematoxylin and eosin--HE). For this study, samples of healthy or lesion skin tissues were obtained from 34 CVL naturally infected dogs classified in three groups: asymptomatic, oligosymptomatic and polisymptomatic. Not only lesion (56.5%) but also healthy skins (31.8%) were positives by IMHC and confirmed by PCR in 97.8% of skin samples. In asymptomatic group, 87.5% dogs were negatives by serological tests, but positives by IMHC in 50% and by PCR in 100%. In oligosymptomatic group, 100%, 85.7% and 28.6% of dogs were positives, respectively by PCR, serological and IMHC tests. In addition, 91.7% of polisymptomatic dogs were serum positive and had intact parasites in the skin. In general, PCR showed higher positivity (100%). The efficiency of each test varied with the evolution of the disease. IMHC may be used to confirm the results of the serology and PCR in inconclusive cases after HE and IMHC. The association of techniques proposed in this study may increase the positivity and contributed to the control of this canine disease.

[Comparative study of diagnostic methods for visceral leishmaniasis in dogs from Ilha Solteira, SP]. / Estudo comparativo dos métodos diagnósticos para Leishmaniose Visceral em cães oriundos de Ilha Solteira, SP.

The purpose of the present work was a comparative study of diagnostic methods for Canine Visceral Leishmaniasis (CVL) using serological methods, enzyme-linked immunosorbent assay (ELISA) and indirect fluorescent antibody test (IFAT), histochemical (HE) and immunohistochemical (IMHC) tests using spleen, lymph node and liver canine tissues. In addition, Polymerase Chain Reaction (PCR) was done in blood and in tissues in order to compare and confirm no conclusive and negative diagnosis by the methods above. For this study, 34 dogs were divided according to clinical signs in asymptomatic, oligosymptomatic and polisymptomatic Leishmania-infected dogs euthanized by Zoonotic Disease Control Center (CCZ) from Ilha Solteira, SP, Brazil. The positivism indexes of ELISA, IMHC, IFAT and HE were 65.0, 62.0, 56.0 and 56.0%, respectively with the highest numbers of positive dogs in polisymptomatic (92.0%) followed by oligosymptomatic (57.0%) and asymptomatic dogs (12.5%). Furthermore, PCR confirmed the positive results and detected DNA in tissues from 100% of negative dogs and 89.0% suspects raising the animal positivism index up to 97.0%. In conclusion, PCR was the most sensitive and a valuable method for a definitive CVL diagnosis.

Estudo comparativo dos métodos diagnósticos para Leishmaniose Visceral em cães oriundos de Ilha Solteira, SP / Comparative study of diagnostic methods for visceral leishmaniasis in dogs from Ilha Solteira, SP

O objetivo da presente pesquisa foi avaliar comparativamente os métodos diagnósticos da Leishmaniose Visceral Canina (LVC), utilizando-se o ensaio imunoenzimático indireto (ELISA), a reação de imunofluorescência indireta (RIFI), a histoquímica (HE) e a imunoistoquímica (IMIQ) em tecidos de órgãos, como o baço, linfonodo e fígado. Além disso, a Reação em Cadeia pela Polimerase (PCR) das amostras de sangue e dos tecidos foi utilizada para comparar e confirmar os diagnósticos negativos e não conclusivos pelos métodos acima. Para esse estudo, foram utilizados 34 cães com diferentes sintomas da LVC, classificados em polissintomáticos, oligossintomáticos e assintomáticos e eutanasiados no Centro de Controle de Zoonoses (CCZ) de Ilha Solteira, SP. Os índices de positividade para os testes ELISA, IMIQ, RIFI e HE foram de 65,0, 62,0, 56,0 e 56,0 por cento, respectivamente, sendo a maior positividade detectada nos cães polissintomáticos (92,0 por cento), seguida pelos oligossintomáticos (57,0 por cento) e assintomáticos (12,5 por cento). A PCR confirmou os resultados positivos pelas outras técnicas e ainda detectou DNA do parasita nos tecidos de 100 por cento dos cães negativos e em 89,0 por cento dos suspeitos, elevando para 97,0 por cento a positividade. Em conclusão, a PCR demonstrou ser o método mais sensível e preciso para o diagnóstico definitivo da LVC.

The purpose of the present work was a comparative study of diagnostic methods for Canine Visceral Leishmaniasis (CVL) using serological methods, enzyme-linked immunosorbent assay (ELISA) and indirect fluorescent antibody test (IFAT), histochemical (HE) and immunohistochemical (IMHC) tests using spleen, lymph node and liver canine tissues. In addition, Polymerase Chain Reaction (PCR) was done in blood and in tissues in order to compare and confirm no conclusive and negative diagnosis by the methods above. For this study, 34 dogs were divided according to clinical signs in asymptomatic, oligosymptomatic and polisymptomatic Leishmania-infected dogs euthanized by Zoonotic Disease Control Center (CCZ) from Ilha Solteira, SP, Brazil. The positivism indexes of ELISA, IMHC, IFAT and HE were 65.0, 62.0, 56.0 and 56.0 percent, respectively with the highest numbers of positive dogs in polisymptomatic (92.0 percent) followed by oligosymptomatic (57.0 percent) and asymptomatic dogs (12.5 percent). Furthermore, PCR confirmed the positive results and detected DNA in tissues from 100 percent of negative dogs and 89.0 percent suspects raising the animal positivism index up to 97.0 percent. In conclusion, PCR was the most sensitive and a valuable method for a definitive CVL diagnosis.

Diagnóstico da Leishmaniose Visceral Canina pelas técnicas de imunoistoquímica e PCR em tecidos cutâneos em associação com a RIFI e ELISA-teste / Canine Visceral Leishmaniasis diagnosis by immunohistochemistry and PCR in skin tissues in association with RIFI and ELISA-test

O objetivo deste trabalho foi avaliar as técnicas de imunoistoquímica (IMIQ) e de PCR (Reação em Cadeia da Polimerase) em tecidos cutâneos para o diagnóstico da Leishmaniose Visceral Canina (LVC) e compará-los com os exames parasitológicos em tecidos corados histoquimicamente (hematoxilina-eosina, HE) e com testes sorológicos, como a Reação de Imunofluorescência Indireta (RIFI) e ensaio imunoenzimático (ELISA). Dos 34 cães naturalmente infectados, classificados em assintomáticos, oligossintomáticos e polissintomáticos, foram coletadas amostras de pele sadia ou com lesão para a realização da IMIQ, HE e PCR. Não somente peles lesionadas (56,5 por cento), mas também sadias (31,8 por cento) encontravam-se positivas pela IMIQ, confirmadas posteriormente pela PCR em 97,8 por cento das amostras. No grupo assintomático, 87,5 por cento estavam negativos pelos testes sorológicos, mas positivos em 50 por cento dos casos pela IMIQ e 100 por cento pela PCR. Entre os oligossintomáticos, 100 por cento, 85,7 por cento e 28,6 por cento encontravam-se positivos, respectivamente, pela PCR, sorologia e IMIQ. Os cães polissintomáticos eram 91,7 por cento soropositivos e tinham parasitas na pele. Em geral, a técnica PCR teve maior positividade (100 por cento). A eficiência dos testes variou de acordo com a evolução da doença, demonstrando a necessidade da associação de técnicas, usando-se IMIQ para confirmação da sorologia e a PCR apenas nos casos suspeitos após a IMIQ. Dessa forma, pode-se aumentar os níveis de positividade e contribuir para o controle desta zoonose.

The purpose of the present study was to evaluate the immunohistochemistry (IMHC) and PCR (Polymerase Chain Reaction) tests for Canine Visceral Leishmaniasis (CVL) diagnosis and compare the results with serological tests such as the indirect fluorescence antibody test (IFAT), ELISA and a parasitological test (microscopic direct examination of the parasite stained with haematoxylin and eosin - HE). For this study, samples of healthy or lesion skin tissues were obtained from 34 CVL naturally infected dogs classified in three groups: asymptomatic, oligosymptomatic and polisymptomatic. Not only lesion (56.5 percent) but also healthy skins (31.8 percent) were positives by IMHC and confirmed by PCR in 97.8 percent of skin samples. In asymptomatic group, 87.5 percent dogs were negatives by serological tests, but positives by IMHC in 50 percent and by PCR in 100 percent. In oligosymptomatic group, 100 percent, 85.7 percent and 28.6 percent of dogs were positives, respectively by PCR, serological and IMHC tests. In addition, 91.7 percent of polisymptomatic dogs were serum positive and had intact parasites in the skin. In general, PCR showed higher positivity (100 percent). The efficiency of each test varied with the evolution of the disease. IMHC may be used to confirm the results of the serology and PCR in inconclusive cases after HE and IMHC. The association of techniques proposed in this study may increase the positivity and contributed to the control of this canine disease.

Diagnóstico da Leishmaniose Visceral Canina pelas técnicas de imunoistoquímica e PCR em tecidos cutâneos em associação com a RIFI e ELISA-teste / Canine Visceral Leishmaniasis diagnosis by immunohistochemistry and PCR

O objetivo deste trabalho foi avaliar as técnicas de imunoistoquímica (IMIQ) e de PCR (Reação em Cadeia da Polimerase) em tecidos cutâneos para o diagnóstico da Leishmaniose Visceral Canina (LVC) e compará-los com os exames parasitológicos em tecidos corados histoquimicamente (hematoxilina-eosina, HE) e com testes sorológicos, como a Reação de Imunofluorescência Indireta (RIFI) e ensaio imunoenzimático (ELISA). Dos 34 cães naturalmente infectados, classificados em assintomáticos, oligossintomáticos e polissintomáticos, foram coletadas amostras de pele sadia ou com lesão para a realização da IMIQ, HE e PCR. Não somente peles lesionadas (56,5%), mas também sadias (31,8%) encontravam-se positivas pela IMIQ, confirmadas posteriormente pela PCR em 97,8% das amostras. Nogrupo assintomático, 87,5% estavam negativos pelos testes sorológicos, mas positivos em 50% dos casos pela IMIQ e 100% pela PCR. Entre os oligossintomáticos, 100%, 85,7% e 28,6% encontravam-se positivos, respectivamente, pela PCR, sorologia e IMIQ. Os cães polissintomáticos eram 91,7% soropositivos e tinham parasitas na pele. Em geral, a técnica PCR teve maior positividade (100%). A eficiência dos testes variou de acordo com a evolução da doença, demonstrando a necessidade da associação de técnicas, usando-se IMIQ para confirmação da sorologia e a PCR apenas nos casos suspeitos apos a IMIQ. Dessa forma, pode-se aumentar os níveis de positividade e contribuir para o controle desta zoonose.

The purpose of the present study was to evaluate the immunohistochemistry (IMHC) and PCR (Polymerase Chain Reaction) tests for Canine Visceral Leishmaniasis (CVL) diagnosis and compare the results with serological tests such as the indirect fluorescence antibody test (IFAT), ELISA and a parasitological test (microscopic direct examination of the parasite stained with haematoxylin and eosin - HE). For this study, samples of healthy or lesion skin tissues were obtained from 34 CVL naturally infected dogs classified in three groups: asymptomatic, oligosymptomatic and polisymptomatic. Not only lesion (56.5%) but also healthy skins (31.8%) were positives by IMHC and confirmed by PCR in 97.8% of skin samples. In asymptomatic group, 87.5% dogs were negatives by serological tests, but positivesby IMHC in 50% and by PCR in 100%. In oligosymptomatic group, 100%, 85.7% and 28.6% of dogs were positives, respectively by PCR, serological and IMHC tests. In addition, 91.7% of polisymptomatic dogs were serum positive and had intact parasites in the skin. In general, PCR showed higher positivity (100%). The efficiency of each test varied with the evolution of the disease. IMHC may be used to confirm the results of the serology and PCR in inconclusive cases after HE and IMHC. The association of techniques proposed in this study may increase the positivity and contributed to the control of this canine disease.