Zoonotic nematode in the city of La Plata, Argentina: Report of a case of Calodium hepaticum in Rattus rattus.

This study describes a case of Calodium hepaticum (Trichinellida: Capillariidae) infection in an adult rat (Rattus rattus) from the periurban area of the city of La Plata in the province of Buenos Aires, Argentina. The rat was found with neurological signs (ataxia, lethargy, and episodes of unresponsiveness) in the food storage of a goat production facility. The liver was observed with hepatomegaly and diffuse and irregular yellowish-white spots appearing in striae or small nodules on the external surface and inside the liver. Subsequent microscopic and histopathological studies were performed. Eggs were observed by direct microscopy of the impression smear of liver tissue. A multifocal granulomatous tissue reaction with different stages of fibrocellular tissue was observed in the liver parenchyma. The granulomas contained adults and degenerated eggs delimited by an intense infiltrate of mononuclear cells. Macro and microscopic observations and histopathological liver lesions were compatible with C. hepaticum infection. To our knowledge, this is the first confirmation of C. hepaticum infection in R. rattus in Argentina, increasing the host record of this parasite and a new record of distribution in goat production systems in the country.

Morphology of the syrinx of three species of birds from Brazilian cerrado (Psittacara leucophthalmus, Rhynchotus rufescens and Cariama cristata): Gross anatomy and light microscopy study.

The aim of this study was to describe the morphology of the trachea and syrinx at macroscopic and light microscopy levels of three species of birds from different orders that inhabit the Brazilian cerrado. For that, five adult specimens (three males and two females of each species) of white-eyed parakeet (Psittacara leucophthalmus), red-winged tinamou (Rhynchotus rufescens) and red-legged seriema (Cariama cristata) were used. The trachea and syrinx of the birds were collected and destined for anatomical and histological studies. The trachea of the studied birds presented an elongated path and originated in the larynx and extended caudally to the syrinx. No sexual dimorphism was observed in the syrinx of the studied species, probably because it is associated with their song, which is very similar between males and females of these species. The findings of this study allowed us to classify the syrinx as tracheal in the white-eyed parakeet and tracheobronchial in the red-winged tinamou and red-legged seriema. In general, the morphological features of the trachea and syrinx were similar to those described for other species of birds, such as the presence of intrinsic and extrinsic syringeal muscles, and the lateral and medial tympaniform membranes, which would represent important anatomical structures in sound production through vibration during expiration and eventual inspiration. The morphological structure of the syrinx in the three avian species of the Brazilian cerrado is consistent with the ability of these avian species to perform a potential vocalization, especially the red-legged seriema that emits characteristic sounds very loud and can carry several kilometres.

Macroscopic and histological aspects of the pharynx and larynx of the giant anteater (Myrmecophaga tridactyla Linnaeus, 1758).

Giant anteater (Myrmecophaga tridactyla) is an endangered species that resides in much of Latin America, but it has been losing its habitat, especially in the Cerrado biome, where it constantly suffers traumas resulting from fires and roadkill. The anatomical knowledge of structures of the respiratory system is important for a better morphophysiological understanding of the species. Thus, this study aimed to perform the macroscopic and histomorphological description of the pharynx and larynx of the giant anteater. Twelve adult giant anteaters were used, three of them fixed in buffered formalin for further dissection and pharynx and larynx macroscopic analysis of structures. From the other animals, samples of the pharynx and larynx were collected and prepared for histological evaluation under optical microscope. Macroscopically, their pharynx and soft palate are extensive, and the anatomical location of these structures and the larynx differs greatly from that described in other species. The larynx, although more caudal, was similar to that of other animals. Histologically, the epithelium of these regions varied between the pseudostratified ciliated columnar and the non-keratinized stratified squamous epithelium. Laryngeal cartilages were composed of elastic (epiglotti) and hyaline cartilages (arytenoid, cricoid and thyroid cartilage), with an ossification process and glandular clusters around the hyaline cartilage. The distinct anatomical location of the pharynx and larynx of Myrmecophaga tridactyla is the main macroscopic finding of this study, besides the length of the pharynx and soft palate of these animals.

Specular microscopy of the corneal endothelial cells of bovines: an ex vivo study.

Background: The endothelium is the most posterior layer of the cornea and is essential for maintaining corneal transparency. Due to variations in corneal endothelial parameters among different species, knowledge of the normal parameters for each species is crucial. Aim: To evaluate the corneal endothelium of bovines using contact specular microscopy. Methods: Twenty eyeballs from 10 male Brangus (Bos taurus) aged 24 months were evaluated. Contact specular microscopy was performed on the central corneal area. The analyzed parameters were endothelial cell density (ECD) and endothelial cell morphology. Results: The ECD in the central area was 1,277 cells/mm2. Regarding the morphology, mainly cells with six (74.3%), five (14.7%) and seven sides (10%) were found. There were no significant differences in ECD and morphology between left and right eyes. Conclusion: Contact specular microscopy facilitated the analysis and measurement of corneal endothelial parameters in bovines. The data obtained will serve as a reference for the analysis of bovine corneal endothelium.

Biochemical changes in the cytoplasm of bovine oocytes during the in vitro maturation process: a Raman microscopy study.

The sequence and chronology of the main biochemical changes occurring in the cytoplasm of bovine oocytes during the in vitro maturation process were tracked by Raman microscopy applied to cells previously subjected to enzymatic digestion of the zona pellucida. Specific spectral markers for proteins, lipids and carbohydrates were used to evaluate the developmental status of the ooplasm at four different times. Spectral changes revealed that lipid accumulation was dominant during the first six hours of culture while protein content reached the average levels characteristic of mature oocytes within the last four hours of the maturation process. A time-dependent decrease in carbohydrates was also observed. Finally, the carbohydrate-to-protein (P1037/P1002) ratio proved to be sensitive enough to determine the cytoplasmic maturation state of bovine oocytes and promises to be useful in future research aimed at optimizing culture conditions through the promotion of protein accumulation in the ooplasm.

Ocular microscopy of Bradypus variegatus.

Bradypus variegatus, popularly known as common sloth or brown-throated sloth, is a species with wide geographical distribution in Brazil. Expand on the literature about the species, we aimed to describe the morphology of its eye and its main annexes. For this study, six frozen specimens donated to the Animal Morphological Research Laboratory of the Federal Rural University of Amazonia were used. The specimens were fixed in a 10% aqueous solution of formaldehyde. The structures that constitute the ocular apparatus were identified and described after dissection and histological analysis using light microscopy. The bulbar conjunctiva presented a stratified cuboidal epithelium, with a considerable number of serous glands. The stroma layer and the posterior limiting lamina were found well defined in the species. The corneoscleral meshwork possessed a large opening for drainage of aqueous humour, followed by a wide retinal region, in which a layer of photoreceptors was visualized. Cones and rods could not be discriminated from each other. Microscopy of the medial angle of the eye revealed the cartilage and glands of the third eyelid, and a well-developed lacrimal gland structure. Bradypus variegatus distinguishing features and similarities in comparison with other domestic and wild mammal and bird species already described were investigated, which expounded the limited knowledge that is presently available on the morphology of wild fauna. The data extend the limited knowledge that are presently available on the ocular histology of B. variegatus.

Prevalência das alterações histopatológicas testiculares encontradas nas raças Curraleiro Pé-Duro, Nelore e seus cruzamentos, criados na região semi-árida do Brasil / Histopathological testicular changes found in the Curraleiro Pé-Duro, Nelore, and its crossings, reared in the semi-arid region of Brazil

O Curraleiro Pé-Duro (CPD) é uma raça nacional, que sofreu um longo processo de seleção natural nas condições do cerrado e semiárido brasileiro. Existem diferenças entre o comportamento reprodutivo das raças naturalizadas brasileiras quando comparadas às raças zebuínas e taurinas. O objetivo deste trabalho foi descrever as alterações histopatológicas testiculares encontradas na raça CPD; Nelore; F1 (CPD x Nelore); Tricross Angus (» CPD + ½ Nelore + ½ Angus); Tricross Senepol (» CPD + ½ Nelore + ½ Senepol), criados na região semiárida do Brasil. Na avaliação microscópica dos 42 testículos, foi observado a ocorrência de 35 casos de degeneração testicular (83,3%), 24 de hipoplasia (57,1%) e 9 de edema (21,4%). Entre os 35 animais que apresentaram diagnóstico de degeneração testicular, 18 casos eram de degeneração leve (51,4%); 7 moderada (20%) e 10 severa (28,6%). Observou-se diversas associações entre degeneração testicular e outro processo patológico no mesmo animal, dentre estes a ocorrência de 17 casos associados à hipoplasia, 5 casos associados a edema e 3 casos associados a hipoplasia e edema no mesmo momento. As alterações histopatológicas testiculares prevalentes nas raças e cruzamentos analisados neste trabalho foram degeneração, hipoplasia e edema, tendo sido a degeneração a de maior acometimento, independentemente da origem racial.(AU)

The Curraleiro Pé-Duro (CPD) is a national breed, which underwent a long process of natural selection in the conditions of the Brazilian cerrado and semi-arid region. There are differences between the reproductive behavior of Brazilian naturalized breeds when compared to zebu and taurine breeds. The objective of this study was to describe the testicular histopathological alterations found in the CPD breed; Nelore; F1 (CPD x Nelore); Tricross Angus (» CPD + ½ Nelore + ½ Angus); Tricross Senepol (» CPD + ½ Nelore + ½ Senepol), bred in the semi-arid region of Brazil. In the microscopic evaluation of the 42 testicles, 35 cases of testicular degeneration (83.3%), 24 cases of hypoplasia (57.1%) and 9 cases of edema (21.4%) were observed. Among the 35 animals that were diagnosed with testicular degeneration, 18 cases were mild degeneration (51.4%); 7 moderate (20%) and 10 severe (28.6%). Several associations were observed between testicular degeneration and another pathological process in the same animal, among which the occurrence of 17 cases associated with hypoplasia, 5 cases associated with edema and 3 cases associated with hypoplasia and edema at the same time. The testicular histopathological alterations prevalent in the breeds and crosses analyzed in this study were degeneration, hypoplasia and edema, with degeneration being the most fruquent, regardless of breed origin.(AU)

Sheep corneal endothelium morphology - evaluation with trypan blue and alizarin red

Background: The endothelium is a layer fundamental to maintaining corneal transparency. In ophthalmology, sheep eyes have been used as a model in research related to corneal transplantation. Different techniques have been used to evaluate the corneal endothelium. Concerning vital dyes, corneal endothelial cell analyses have not yet been studied in ovines. The purpose of the present study was to evaluate the morphology of endothelial cells from different regions of the cornea of sheep after staining with alizarin red and trypan blue using an optical microscope. Materials, Methods & Results: Twenty healthy eyes of 10 male sheep obtained from a licensed commercial slaughterhouse were studied. The study was approved by the Research Committee of the Faculty of Veterinary at UFRGS and followed the ethical standards of the Association for Research in Vision and Ophthalmology (ARVO). Immediately after the slaughter, the eyes were enucleated and underwent eye examination. The corneal endothelium was stained with trypan blue and alizarin red and examined and photographed using an optical microscope. The central, superior, inferior, nasal and temporal areas of the cornea were evaluated for cell morphology. Data were compared by t-tests. Differences were considered statistically significant at P < 0.05. Immediately after staining the corneal endothelium, it was possible to examine with an optical microscope, obtain images and analyse the shape of endothelial cells from all regions of the sheep cornea. Polygonal, uniform and continuous cells were observed in all samples studied. Considering all the corneas analysed, cells with 6 sides (75.11%), 5 sides (12.76%) and 4 sides (12.12%) were found. In the central region of the cornea 75.91% of cells with 6 sides, 12.6% of cells with 5 sides and 11.48% with 7 sides were found. In the superior region of the cornea 76.07% of cells with 6 sides, 13.25% with 5 sides and 10.68% with 7 sides were found. In the lower region were found 74.72% of cells with 6 sides, 13% with 5 sides and 12.27% with 7 sides. In the temporal region, 74.14% were 6-sided cells, 11.42% had 5 sides, and 14.43% had 7 sides. Furthermore, in the nasal region, 74.72% of the cells had 6 sides, 13.54% had 5 sides, and 11.73% had 7 sides. No significant differences were found between cell morphology in all corneal regions evaluated. In addition, no significant difference was found when comparing the right eye with the left eye. Discussion: Different methods are used for the analysis of corneal endothelium. For ex vivo research optical microscopy after endothelial staining is an alternative low-cost technique that allows the analysis of all regions of the cornea. Quantitative analyses must characterise the endothelial parameters of the different species. The analysis of the morphology of corneal endothelium with an optic microscope after staining with alizarin red has been described as an effective, rapid and cost-efficient method, since this dye blends with the borated cells, allowing identification. In the present study, using optical microscopy and coloration with alizarin red it was possible to explore and obtain images of the ovine endothelium of all regions of the cornea. In the current study, the endothelium had a predominance of cells will 6 sides in all regions studied. This study allowed us to obtain images of the endothelium as well as quantitative data on the morphology of the different regions of the sheep cornea. This study demonstrated that morphology did not differ between the central and peripheral regions. The findings of this study represent a further source of reproducible data that should be considered when using sheep cornea as ex vivo model for experimental research.

Comparison of Liver Condemnation and Bile Microscopy As Tools to Estimate Fasciola hepatica Prevalence and Burden in the Anta Province of Cusco in Peru.

Fascioliasis is a zoonotic infection linked to significant economic losses in the livestock industry. Infection prevalence and estimated financial burden vary across locations owing to different diagnostic tests used. The accuracy of liver condemnation to estimate the prevalence and costs of fascioliasis has seldom been evaluated. We performed a pilot study to determine the prevalence and burden of Fasciola hepatica infection among cattle slaughtered at the municipal abattoir in the Anta province of the Cusco highlands in Peru. We compared liver condemnation with bile microscopy for the diagnosis of infection and prediction of carcass weight. Data were collected from 2009 slaughtered cattle for 1 year. The overall prevalence of Fasciola infection by bile microscopy was 62.5% (1247/2009). A higher prevalence was observed after the rainy season from March to August than from September to February (p < 0.01). Fascioliasis prevalence during the first 6 months was 77.4% (714/923), combining the results of condemnation and microscopy. Bile microscopy diagnosed more infections than liver condemnation (62.7% (579/923) versus 55.4% (511/923), McNemar test p < 0.01). The agreement of the bile microscopy testing with liver condemnation was fair (κ = 0.247). Animal age, gender, breed, and liver condemnation predicted carcass weight [F (df 4, 704) = 61.1, p < 0.001]. Liver condemnation and bile microscopy are complementary tools for evaluation of the prevalence and burden of fascioliasis in livestock. Large scale studies are warranted to confirm our results.

Optical imaging of cleavage stage bovine embryos using hyperspectral and confocal approaches reveals metabolic differences between on-time and fast-developing embryos.

The assessment of embryo quality aims to enhance subsequent pregnancy and live birth outcomes. Metabolic analysis of embryos has immense potential in this regard. As a step towards this goal, here we assess the metabolism of bovine embryos using label-free optical imaging. We compared embryos defined as either on-time or fast-developing, as fast dividing embryos are more likely to develop to the blastocyst stage. Specifically, bovine embryos at 48 (Day 2) and 96 (Day 4) hours post fertilization were fixed and separated based on morphological assessment: on-time (Day 2: 2 cell; Day 4: 5-7 cell) or fast-developing (Day 2: 3-7 cell; Day 4: 8-16 cell). Embryos with different developmental rates on Day 2 and Day 4 were correlated with metabolic activity and DNA damage. Confocal microscopy was used to assess metabolic activity by quantification of cellular autofluorescence specific for the endogenous fluorophores NAD(P)H and FAD with a subsequent calculation of the optical redox ratio. Separately, hyperspectral microscopy was employed to assess a broader range of endogenous fluorophores. DNA damage was determined using γH2AX immunohistochemistry. Hyperspectral imaging showed significantly lower abundance of endogenous fluorophores in fast-developing compared to on-time embryos on Day 2, indicating a lower metabolic activity. On Day 4 of development there was no difference in the abundance of FAD between on-time and fast-developing embryos. There was, however, significantly higher levels of NAD(P)H in fast-developing embryos leading to a significantly lower optical redox ratio when compared to on-time embryos. Collectively, these results demonstrate that fast-developing embryos present a 'quiet' metabolic pattern on Day 2 and Day 4 of development, compared to on-time embryos. There was no difference in the level of DNA damage between on-time and fast-developing embryos on either day of development. To our knowledge, this is the first collective use of confocal and hyperspectral imaging in cleavage-stage bovine embryos in the absence of fluorescent tags.

Evaluation of stallion sperm motility with ImageJ using a cell phone camera and a light microscope / [Avaliação da motilidade espermática de garanhões com ImageJ usando-se câmera de celular e microscópio óptico]

This study aimed to determine the accuracy of assessing stallion sperm motility using a light microscope, a cell phone camera, and a free computer-assisted semen analysis (FCASA) package for ImageJ. The total motility of frozen (n=22) and cooled (n=48) equine semen was determined by FCASA and compared to the results of subjective visual analysis (SVA) by two technicians. Frozen samples were also evaluated by a commercial computer-assisted semen analysis (CCASA) system. The Friedman test revealed no significant differences (P>0.05) between cooled samples analyzed by FCASA (38.0) and SVA (technician 1, 40.0; technician 2, 40.0), nor between frozen samples analyzed by FCASA (23.36 ± 15.9), SVA (25.5 ± 18.8 and 25.8 ± 18.5), and CCASA (25.2 ± 18.3). However, mean FCASA results were underestimated by 7.2% compared with CCASA. The correlation between FCASA and CCASA was significant and strong (P<0.0001, r=0.95). Chi-squared tests indicated that FCASA provided similar results (P=0.14) to the reference method (CCASA), but SVA had lower accuracy (P=0.04). ImageJ analysis of cell phone videos captured under a light microscope can be used for estimation of stallion sperm motility with comparable accuracy to commercial systems.(AU)

O objetivo deste estudo foi testar as configurações necessárias para avaliar a motilidade espermática total de garanhões, mediante o uso de ImageJ, microscópio óptico e câmera de celular. Os valores de motilidade total das amostras de sêmen equino congeladas (22) e refrigeradas (48) foram comparados por análise visual (SVA) e pelo plugin do ImageJ (CASAF). Amostras congeladas também foram comparadas por um CASA comercial (CCASA). O teste de Friedman não resultou em diferença estatística (P>0,05) entre as 48 amostras analisadas com CASAF (38,0) e SVA de dois avaliadores (40,0 e 40,0). A comparação das 22 amostras congeladas entre CASAF (23,36±15,9), SVA (25,5±18,8 e 25,8±18,5) e CCASA (25,2±18,3) também não resultou em diferença estatística, sendo que a média dos resultados obtidos com CASAF subestimou a obtida com o CCASA em 7,2%. A correlação entre CASAF e CCASA foi significativamente elevada (r=0,95, P<0,0001). O teste de qui-quadrado resultou em proporção de acertos semelhantes entre o CASAF e o CCASA (P=0,14), enquanto SVA resultou em proporção diferente (P=0,04), indicando menor acurácia. O uso de microscópio óptico e câmera de celular foi útil para obter vídeos de sêmen de garanhões a serem analisados com ImageJ, proporcionando resultados de motilidade total equiparáveis a sistemas comerciais.(AU)

An integrative approach assesses the intraspecific variations of Procamallanus (Spirocamallanus) inopinatus, a common parasite in Neotropical freshwater fishes, and the phylogenetic patterns of Camallanidae.

Integrative taxonomy was used to evaluate two component populations of Procamallanus (Spirocamallanus) inopinatus in Brazil and the phylogeny Camallanidae. Parasite populations were collected in the characiform Anostomoides passionis from River Xingu (Amazon basin) and Megaleporinus elongatus from River Miranda (Paraguay basin). Morphology was analysed using light and scanning electron microscopy (SEM). Genetic characterization was based on partial sequences of the 18S and 28S rDNA, and COI mtDNA. Phylogenies were based on 18S and COI due to data availability. Generalized Mixed Yule Coalescent (GMYC), Poisson Tree Process (PTP) and *BEAST were used for species delimitation and validation. SEM revealed for the first time the presence of minute denticles and pore-like structures surrounding the oral opening, phasmids in females and confirmed other important morphological aspects. Statistical comparison between the two-component populations indicated morphometric variations, especially among males. The different component population of P. (S.) inopinatus showed variable morphometry, but uniform morphology and were validated as conspecific by the GMYC, PTP and *BEAST. Some camallanid sequences in GenBank have incorrect taxonomic labelling. Host, environment and geographic aspects seem to be related to some lineages within Camallanidae; however, their real phylogenetic meanings are still unclear.

V-Prechop nucleodissection technique feasibility in the phacoemulsification in dogs with cataracts. Clinical aspects and specular microscopy / Exequibilidade da técnica V-Prechop de nucleodissecção na facoemulficação em cães com catarata. Aspectos clínicos e microscopia especular

The aim of this study was to evaluate the feasibility of the V-Prechop nucleodissection technique in the phacoemulsification in dogs and the clinical aspects and of the specular microscopy in the postoperative period. Fourty three dogs of different breeds, males and females, aged 3 to 10 years, with mature (n=22) and immature (n=21) cataracts were used. After surgery, patients were evaluated weekly for visual acuity, intraocular pressure, and endothelial corneal cell density (by non-contact specular microscopy) and quantitative flare (by laser flare photometry) during different periods. The "V-prechop" technique presented technical difficulties in implementation in patients with a mature cataract. In selected cases of patients with an immature cataract, the technique can be employed, as the nuclei are softer, allowing confection of the linear fragments in "V" to be performed. In addition, the eyes of dogs with a mature cataract presented more intensive postoperative uveitis, probably related to the greater difficulty inconducting the "V" nucleodissection. There was decreased endothelial corneal cell density in dogs with mature and immature cataracts. This occurrence was greater in patients with a mature cataract, given the increased intraocular manipulation and surgical time due to the difficulty in performing the "V" nucleodissection. According to the results obtained, the "V-prechop" nucleodissection technique can be indicated in selective cases of dogs with an immature cataract.(AU)

O objetivo deste estudo foi avaliar a viabilidade da técnica V-Prechop de nucleodissecção na facoemulsificação em cães e os aspectos clínicos e de microscopia especular no período pós-operatório. Quarenta e três cães de diferentes raças foram utilizados, machos e fêmeas, com idade entre três a dez anos e portadores de cataratas maduras (n = 22) e imaturas (n = 21). Após a cirurgia, os pacientes foram avaliados semanalmente quanto à acuidade visual, pressão intraocular e densidade celular endotelial da córnea (por microscopia especular sem contato) e quantitativo de flare (por flarefotometria a laser), em diferentes períodos. A técnica "V-prechop" revelou dificuldades técnicas em pacientes com catarata madura. Em casos selecionados de pacientes com uma catarata imatura, a técnica pode ser empregada, pois os núcleos são menos densos, o que permitiu a confecção dos fragmentos lineares em "V". Além disso, os olhos de cães com uma catarata madura apresentaram uveíte posoperatória mais intensa, provavelmente relacionada à maior dificuldade em conduzir a confecção das fraturas em "V". Houve diminuição da densidade das células corneais endoteliais cães com catarata madura e imatura. Essa ocorrência foi mais pronunciada em pacientes com uma catarata madura, devido a maior manipulação cirúrgica intraocular e tempo cirúrgico, pois houve maior dificuldade técnica em realizar a confecção dos fragmentos em "V". De acordo com os resultados obtidos, a técnica de nucleodissecção "V-prechop" pode ser indicada em casos seleconados de cães com catarata imatura.(AU)

Macro and microscopic comparison of the upper pole of the spleen with the lower pole in partial splenectomy

Purpose To evaluate the viability of the upper (UP) and lower pole (LP) of the spleen from a macro and microscopic point of view, after subtotal splenectomy with preservation (SSP) of the UP and the LP. Methods Seventeen male Wistar rats, two months old, were submitted to SSPUP and SSPLP and 5 to simulated operation (SG). After 80 days, the rats were euthanized, and the remaining LP and UP and intact spleens were evaluated macroscopically and microscopically. Results Two rats died during the operation. Macroscopic analysis showed that in 15 LP, one of them was not viable and in 15 UP and in 5 spleens in the SG, all were viable. In the statistical analysis, there was no difference in relation to viability. The LP and UP analyzed showed variation. As for the length, the UP increased significantly; however, in relation to the width, there was a significant increase in the LP in relation to the UP. In addition, the weight of the UP was significantly greater than that of the LP. Microscopic analysis attested viability of the splenic remnants. Conclusion There was no significant difference regarding the viability of UP and LP, in macroscopy and microscopy.(AU)

V-Prechop nucleodissection technique feasibility in the phacoemulsification in dogs with cataracts. Clinical aspects and specular microscopy / Exequibilidade da técnica V-Prechop de nucleodissecção na facoemulficação em cães com catarata. Aspectos clínicos e microscopia especular

The aim of this study was to evaluate the feasibility of the V-Prechop nucleodissection technique in the phacoemulsification in dogs and the clinical aspects and of the specular microscopy in the postoperative period. Fourty three dogs of different breeds, males and females, aged 3 to 10 years, with mature (n=22) and immature (n=21) cataracts were used. After surgery, patients were evaluated weekly for visual acuity, intraocular pressure, and endothelial corneal cell density (by non-contact specular microscopy) and quantitative flare (by laser flare photometry) during different periods. The "V-prechop" technique presented technical difficulties in implementation in patients with a mature cataract. In selected cases of patients with an immature cataract, the technique can be employed, as the nuclei are softer, allowing confection of the linear fragments in "V" to be performed. In addition, the eyes of dogs with a mature cataract presented more intensive postoperative uveitis, probably related to the greater difficulty inconducting the "V" nucleodissection. There was decreased endothelial corneal cell density in dogs with mature and immature cataracts. This occurrence was greater in patients with a mature cataract, given the increased intraocular manipulation and surgical time due to the difficulty in performing the "V" nucleodissection. According to the results obtained, the "V-prechop" nucleodissection technique can be indicated in selective cases of dogs with an immature cataract.

O objetivo deste estudo foi avaliar a viabilidade da técnica V-Prechop de nucleodissecção na facoemulsificação em cães e os aspectos clínicos e de microscopia especular no período pós-operatório. Quarenta e três cães de diferentes raças foram utilizados, machos e fêmeas, com idade entre três a dez anos e portadores de cataratas maduras (n = 22) e imaturas (n = 21). Após a cirurgia, os pacientes foram avaliados semanalmente quanto à acuidade visual, pressão intraocular e densidade celular endotelial da córnea (por microscopia especular sem contato) e quantitativo de flare (por flarefotometria a laser), em diferentes períodos. A técnica "V-prechop" revelou dificuldades técnicas em pacientes com catarata madura. Em casos selecionados de pacientes com uma catarata imatura, a técnica pode ser empregada, pois os núcleos são menos densos, o que permitiu a confecção dos fragmentos lineares em "V". Além disso, os olhos de cães com uma catarata madura apresentaram uveíte posoperatória mais intensa, provavelmente relacionada à maior dificuldade em conduzir a confecção das fraturas em "V". Houve diminuição da densidade das células corneais endoteliais cães com catarata madura e imatura. Essa ocorrência foi mais pronunciada em pacientes com uma catarata madura, devido a maior manipulação cirúrgica intraocular e tempo cirúrgico, pois houve maior dificuldade técnica em realizar a confecção dos fragmentos em "V". De acordo com os resultados obtidos, a técnica de nucleodissecção "V-prechop" pode ser indicada em casos seleconados de cães com catarata imatura.

Methods for glomerular quantification in dogs: a comparative study / Metodologias para quantificação glomerular em cães: um estudo comparativo

It was evaluated the different methodologies for glomerular enumeration described in the literature in a quantitative, double-blind study on the number of glomeruli present in kidney biopsy specimens obtained using a Tru-cut-type biopsy needle. Eighteen samples were taken (n=18) from various regions of canine kidneys harvested immediately after euthanasia. Sample collection was carried out using Tru-cut-type, semi-automated, 16 gauge needles. Reading and evaluation of the specimens were performed independently by four researchers by means of eye loupe inspection, light microscopy, light microscopy with a reduced condenser aperture, and dissection microscopy. Samples were also subjected to histopathological evaluation for comparison purposes. There was no inter-researcher variation for any of the tested methods. The glomerular counts obtained using light microscopy with reduced condenser aperture were similar to those reported in the histopathological evaluation (P=0.44);whereas, the other glomerular enumeration methods yielded significantly lower counts when compared to the histopathological analysis (eye loupe inspection: P<0.01; conventional light microscopy: P=0.02; dissection microscopy: P<0.01).Light microscope with lowering of the condenser lens delivers better results than conventional light microscopy and eye loupe inspection. Results obtained using dissection microscopy and eye loupe inspection did not correlate well with those provided by histopathological evaluation; consequently, these two methods should be avoided.(AU)

Avaliou-se as diferentes metodologias de quantificação glomerular descritas na literatura em um estudo quantitativo, duplo-cego, sobre o número de glomérulos presentes em amostras de biópsias renais obtidas com agulha de biópsia tipo Tru-cut. Dezoito amostras foram coletadas (n=18) de várias regiões de rins caninos colhidos imediatamente após a eutanásia. A coleta das amostras foi realizada com agulhas semi-automáticas de calibre 16, tipo Tru-cut. A leitura e avaliação das amostras foram realizadas de forma independente por quatro pesquisadores por meio de inspeção visual, microscopia de luz, microscopia de luz com redução da lente condensadora e microscopia de dissecção. As amostras também foram submetidas à avaliação histopatológica para fins de comparação e variação do pesquisador para qualquer um dos métodos testados. As contagens glomerulares obtidas por microscopia de luz com redução da lente condensadora foram semelhantes àquelas encontradas na avaliação histopatológica (P=0.44), enquanto que os outros métodos de quantificação glomerular produziram contagens significativamente mais baixas a análise histopatológica (inspeção da lupa dos olhos: P<0.01; microscopia de luz convencional: P=0.02; microscopia dissecação: P<0.01). Microscópio de luz com abaixamento da lente condensadora fornece melhores resultados do que microscopia de luz convencional e inspeção da lupa. As contagens obtidas usando microscopia de dissecção e inspeção de lupa de olho não se correlacionaram bem com aqueles fornecidos pela avaliação histopatológica; consequentemente, estes dois métodos devem ser evitados.(AU)

Microscopic alterations in Fasciola hepatica from sheep treated with albendazole / Alterações microscópicas em Fasciola hepatica de ovelhas tratadas com albendazol

Currently, albendazole is one of the most commonly used drugs because of its affordability. The objective was to evaluate the histopathology of Fasciola hepatica specimens. For this, the efficacy test was performed on sheep treated with albendazole at the dose recommended for F. hepatica, in which the helminths recovered at necropsy were counted and separated for histology. Spermatogenic cells from parasites recovered from treated and control sheep were examined by microscopy. The fecal egg-count reduction test revealed 97.06% efficacy of albendazole in the treatment of F. hepatica. Changes in testicular tubule cells started 48 hours after treatment and became evident within 72 hours, at which point it became difficult to identify cell types. Primary and secondary spermatogonia became increasingly rare and intercellular vacuolization was more evident. Signs of apoptosis, with pycnotic nuclei and evidence of keriorrexia were observed at all times. Cell debris was identified 96 hours after treatment. The results indicated that parasitic spermatogenesis was severely affected by albendazole and demonstrated the importance of the use of histopathology for the diagnosis of therapeutic efficacy in field strains.(AU)

Na atualidade, o albendazol é uma das drogas mais usadas devido à sua acessibilidade econômica. O objetivo foi avaliar a histopatologia dos espécimes de Fasciola hepatica. Para isso, foi realizado o teste de eficácia em ovinos tratados com albendazol na dose recomendada para Fasciola hepatica, no qual os helmintos recuperados em necropsia foram contabilizados e separados para histologia. As células espermatogênicas de parasitas recuperados de ovinos tratados e controle foram examinadas por microscopia. O teste de redução de ovos por grama de fezes revelou 97,06% de eficácia do albendazol no tratamento de F. hepatica. As alterações nas células dos túbulos testiculares iniciaram-se 48 horas após o tratamento e tornaram-se evidentes em 72 horas, altura em que tornou-se difícil identificar os tipos de células. As espermatogônias primárias e secundárias tornaram-se cada vez mais raras e a vacuolização intercelular foi mais evidente. Sinais de apoptose, com núcleos picnóticos e evidência de cariorrexia foram observados em todos os momentos. Os detritos celulares foram identificados 96 horas após o tratamento. Os resultados indicaram que a espermatogênese parasitária foi severamente afetada pelo albendazol e demonstrou a importância do uso da histopatologia para o diagnóstico de eficácia terapêutica em cepas de campo.(AU)

Morphology of endothelial cells from different regions of the swine cornea

Background: The corneal endothelium is a monolayer of polygonal cells which constitute the last layer of the cornea. The integrity of this layer is critical to cornea transparency. The characterization of normal corneal endothelial morphology is important not only to clinical evaluation but also to selection of areas of the cornea with better quality to be employed as donor tissue. The aim of the present study was to evaluate the morphology of endothelial cells from different regions of the swine cornea after alizarin red staining using optical microscopy.Materials, Methods & Results: Twenty-four healthy eyes from 12 swine Large White breed, with 14-monthold, males or females obtained from a licensed Brazilian commercial slaughterhouse were studied. Immediately after humane slaughter, the eyes were enucleated and submitted to ophthalmic examination. Eyes with signs of diseases of the anterior segment were excluded. The cornea, with 3 mm of the sclera, was removed and placed on a glass microscope slide with the endothelial side up. Four radial incisions were made in the periphery of the cornea to better accommodate the cornea on the microscope slide. Alizarin red was diluted in isotonic solution (0.2 g/100 mL) and the pH was adjusted to 4.2 with hydrochloric acid. Three drops of alizarin red were placed on the corneal endothelium. After 90 s, the dye was removed from the cornea with balanced saline solution. The corneal endothelium was examined and photographed using an optical microscope. All evaluations were performed by the same investigator. Photomicrographs were taken of central, superior, inferior, nasal and temporal corneal areas. Parameter studied included endothelial cell morphology. For the statistical analysis, was employed the ANOVA variance test (repeated measures). Differences were considered statistically significant at P < 0.05.[...]