Virulence Profiles of Wild-Type, P.1 and Delta SARS-CoV-2 Variants in K18-hACE2 Transgenic Mice.

Since December 2019, the world has been experiencing the COVID-19 pandemic caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), and we now face the emergence of several variants. We aimed to assess the differences between the wild-type (Wt) (Wuhan) strain and the P.1 (Gamma) and Delta variants using infected K18-hACE2 mice. The clinical manifestations, behavior, virus load, pulmonary capacity, and histopathological alterations were analyzed. The P.1-infected mice showed weight loss and more severe clinical manifestations of COVID-19 than the Wt and Delta-infected mice. The respiratory capacity was reduced in the P.1-infected mice compared to the other groups. Pulmonary histological findings demonstrated that a more aggressive disease was generated by the P.1 and Delta variants compared to the Wt strain of the virus. The quantification of the SARS-CoV-2 viral copies varied greatly among the infected mice although it was higher in P.1-infected mice on the day of death. Our data revealed that K18-hACE2 mice infected with the P.1 variant develop a more severe infectious disease than those infected with the other variants, despite the significant heterogeneity among the mice.

Detection of Trypanosoma vivax in tissues of experimentally infected goats: what is the role of adipose tissue in the life cycle of this protozoon?

Trypanosomiasis, caused by Trypanosoma vivax, is responsible for great economic losses among livestock in Africa and South America. During the life cycle of these parasites, they may present different morphological, metabolic and physiological characteristics depending on the interactions that are encountered at each point of their life cycle. Although T. vivax is frequently reported in the circulation of its mammalian hosts, it has the ability to migrate to the tissues of these individuals. However, this characteristic is poorly understood. In this context, we aimed to investigate the presence of T. vivax and the changes caused in different tissues of experimentally infected goats. Despite the animals were not perfused before tissues collection, using different approaches, we demonstrated its presence in different samples, including in the adipose tissue and skin of infected animals. In addition, a mononuclear inflammatory reaction, mostly characterized by an infiltrate of lymphocytes, plasma cells and macrophages were observed. The results highlight the possibility that, like other trypanosomatids, T. vivax may use these tissues during its life cycle. Future studies aiming to elucidate the length of time for which T. vivax remains active in these sites, and whether it uses these sites as a refuge from trypanocidal drugs, and whether it is capable of recolonizing the blood circulation, are much needed.

Nellore cloned calf viability during the postnatal period / Viabilidade de bezerros nelores clonados durante o período pós natal imediato

ABSTRACT: This study evaluated the viability of Nellore cloned calves derived from somatic cell nuclear transfer (SCNT) and compare their viability with animals of the same breed derived from in vitro fertilization (IVF). Thus, two groups were formed. Group I (GI) consisted of 10 calves derived from SCNT and group II (GII) consisted of 10 calves derived from IVF. The differences detected between the groups were in the physical examination of the respiratory tract in GI, which represented the most common clinical-pathological disturbances. The Apgar index score indicated that 80% of GI animals were depressed and all had pale mucous membranes. Thus, anemia was reported in GI. In GII, this started at 12 h of life and was probably caused by an iron deficiency. Moreover, total calcium and ionized calcium levels were higher in GI immediately after birth. These alterations probably resulted in a high incidence of mortality in GI, reaching 90% of the calves, whereas mortality was only 20% for the calves in GII. In conclusion, cloned calves, which were derived from SCNT, had physiological and metabolic alterations after delivery, leading to a higher mortality rate during the perinatal period.

RESUMO: O objetivo deste trabalho foi avaliar a viabilidade de bezerros da raça Nelore oriundos da técnica da transferência nuclear de células somáticas (TNCS), no período pós natal imediato, comparando-a com animais desta mesma raça, oriundos de fertilização in vitro (FIV). Para tanto, os animais foram alocados em dois grupos, a saber: Grupo I (GI) - 10 animais frutos de TNCS; e, Grupo II (GII) - 10 animais oriundos de FIV. Nos respectivos bezerros, todos obtidos por cesariana, foram realizadas as avaliações físicas, escore de APGAR, bem como coleta de amostras de sangue nos momentos 0 (ao nascimento), às 2, 4, 6 e 12 horas de vida, a fim de avaliar os resultados de eritrograma, análises bioquímicas e hormonais, comparando-os entre os grupos e momentos. Nos animais que vieram a óbito foi realizada a necropsia para investigar a causa mortis. As diferenças observadas foram em relação aos achados clínico-patológicos, envolvendo, principalmente, o sistema respiratório caracterizado por bradpneia associada à dispneia, e a presença de edema e atelectasia pulmonar observadas no GI. Ademais, após a colostragem notou-se que 80% dos animais avaliados não foram capazes de manter a glicemia sendo mais evidentes nos animais do GII, possivelmente devido à hiperinsulinemia que se manifestou neste grupo ao longo de todo o período experimental. A anemia após o nascimento foi evidente no grupo de bezerros clonados ao longo de todo período de avaliação ao contrário dos bezerros oriundos de GII, a qual foi observada somente às 12 horas de vida, sendo, possivelmente, de origem ferropriva. Casos de hipercalcemia foram denotados nos animais do GI ao nascimento, sendo possivelmente associados à asfixia perinatal. Estas alterações, em conjunto, levaram a uma taxa de mortalidade de 90% dos animais do GI e de 20% dos bezerros do GII. Conclui-se que os animais oriundos da TNCS apresentam alterações fisiológicas e metabólicas após o nascimento, responsáveis, em grande parte, pela maior taxa de óbitos dentro do período perinatal.

Detection of Trypanosoma vivax in tissues of experimentally infected goats: what is the role of adipose tissue in the life cycle of this protozoon? / Detecção de Trypanosoma vivax em tecidos de caprinos experimentalmente infectados: qual é o papel do tecido adiposo no ciclo de vida desse protozoário?

Abstract Trypanosomiasis, caused by Trypanosoma vivax, is responsible for great economic losses among livestock in Africa and South America. During the life cycle of these parasites, they may present different morphological, metabolic and physiological characteristics depending on the interactions that are encountered at each point of their life cycle. Although T. vivax is frequently reported in the circulation of its mammalian hosts, it has the ability to migrate to the tissues of these individuals. However, this characteristic is poorly understood. In this context, we aimed to investigate the presence of T. vivax and the changes caused in different tissues of experimentally infected goats. Despite the animals were not perfused before tissues collection, using different approaches, we demonstrated its presence in different samples, including in the adipose tissue and skin of infected animals. In addition, a mononuclear inflammatory reaction, mostly characterized by an infiltrate of lymphocytes, plasma cells and macrophages were observed. The results highlight the possibility that, like other trypanosomatids, T. vivax may use these tissues during its life cycle. Future studies aiming to elucidate the length of time for which T. vivax remains active in these sites, and whether it uses these sites as a refuge from trypanocidal drugs, and whether it is capable of recolonizing the blood circulation, are much needed.

Resumo A tripanossomíase, causada por Trypanosoma vivax, é responsável por grandes perdas econômicas na bovinocultura da África e da América do Sul. Durante seu ciclo de vida, o parasita pode apresentar diferentes características morfológicas, metabólicas e fisiológicas em função das interações que ele encontra em cada ponto do seu ciclo. Embora o T. vivax seja reportado, frequentemente, na circulação dos seus hospedeiros mamíferos, o protozoário tem a capacidade de migrar para os tecidos desses indivíduos. Entretanto, essa característica é pobremente conhecida. Neste contexto, o objetivo foi verificar a presença, assim como as alterações causadas pelo T. vivax nos diferentes tecidos de caprinos experimentalmente infectados. Apesar dos animais não terem sido perfundidos antes da coleta dos tecidos, utilizando-se diferentes abordagens, foi evidenciada a presença do T. vivax em diferentes amostras teciduais, incluindo no tecido adiposo e pele dos animais infectados. Além disso, foi observada reação inflamatória mononuclear, caracterizada majoritariamente por infiltrado de linfócitos, plasmócitos e macrófagos. Os resultados evidenciam a possibilidade de que, assim como outros tripanossomatídeos, T. vivax pode usar esses tecidos durante o seu ciclo de vida. São necessários futuros estudos, objetivando elucidar o período em que o T. vivax permanece ativo nesses sítios, se ele utiliza esses locais como refúgio das drogas tripanocidas, e se ele é capaz de recolonizar a circulação sanguínea.

Eosinophilic inflammation in lymph nodes of dogs with visceral leishmaniasis.

Eosinophils are traditionally associated with the immune response against helminth parasites. However, several studies have demonstrated that these cells have a role regarding protective immunity in leishmaniasis. Here, we examined the relationship between the presence of eosinophils and parasite load in biopsy samples from dogs, obtained through fine needle puncture and aspiration of lymph nodes. Histological slides containing biopsy material from lymph nodes of dogs with canine visceral leishmaniasis and healthy dogs were used to obtain baseline eosinophil counts. Subsequently, scrapings were taken from slides for DNA extraction and determination of parasite load, using real-time PCR (qRT-PCR). Additionally, production of nitric oxide (NO) and reactive oxygen species (ROS) levels by eosinophils in the peripheral blood of dogs with canine visceral leishmaniasis and healthy dogs was measured. The eosinophil percentage were higher in lymph nodes of infected group, and the parasite load showed a significant negative correlation with the eosinophil count. The production of NO and ROS by eosinophils in the peripheral blood was higher in the dogs with canine visceral leishmaniasis. All the results together suggest that eosinophils may participate in antileishmanial immunity in canine visceral leishmaniasis.

Congenital Toxoplasmosis in Chronically Infected and Subsequently Challenged Ewes.

This experiment studied congenital transmission in sheep experimentally infected with oocysts of Toxoplasma gondii and reinfected at one of three stages of pregnancy. Twenty ewes were experimentally infected with T. gondii strain ME49 (day 0). After the T. gondii infection became chronic (IFAT≤512), the ewes were allocated with rams for coverage. After the diagnosis of pregnancy, these ewes were allocated into four experimental groups (n = 5): I-reinfected with T. gondii on the 40th day of gestation (DG); II-reinfected on DG 80; III-reinfected on DG 120; and IV-saline solution on DG 120 (not reinfected). Five ewes (IFAT<64) were kept as negative controls (uninfected, group V), therefore in groups I-III were infected prior to pregnancy and re-infected during pregnancy, group IV was only infected prior to pregnancy, and group V was not infected. Parasitism by T. gondii was investigated (histopathology, immunohistochemistry, mouse bioassay and PCR) in mothers and lambs tissue. All ewes produced lambs serologically positive for T. gondii. The results of the mouse bioassay, immunohistochemistry and PCR assays revealed the presence of T. gondii in all 20 sheep and their lambs. The congenital transmission of T. gondii was associated with fetal loss and abnormalities in persistently infected sheep and in ewes infected and subsequently reinfected by this protozoan. Therefore, congenital T. gondii infection was common when ewes were chronically infected prior to pregnancy, with or without reinfection during at various stages of gestation.

Beta therapy with 90strontium as single modality therapy for canine squamous cell carcinoma in third eyelid / Betaterapia com estrôncio-90 como modalidade única de terapia para o carcinoma de células escamosas na terceira pálpebra em cães

The purpose was to evaluate the effectiveness of beta-radiation with strontium-90 as single modality treatment of canine third eyelid squamous cell carcinoma (SCC). Nine dogs diagnosed with third eyelid SCC were treated with strontium-90. Radiation therapy was administered in four fractions of 100cGy per site every four days and at a depth of 0.2cm (Strontium-90 build' up) in each fraction. Radiation with beta therapy was well tolerated in all animals with no occurrence of radiation induced cataracts. In all cases, there were increased signs of conjunctival inflammation around the mass, which subsided with topical anti-inflammatory. Two dogs required surgical treatment for local tumor recurrence at 150 days and 352 days. In the remaining seven cases, disease free interval ranged from 1239 days to 2555 days. Beta therapy using 90Sr may be a valid alternative for the treatment of third eyelid SCC in dogs.

O objetivo foi avaliar a eficácia da betaterapia com estrôncio-90 como modalidade única de tratamento em carcinomas de células escamosas (CEC) da terceira pálpebra de cães. Nove cães foram diagnosticados com CEC de terceira pálpebra, que foram tratados com estrôncio-90. A radioterapia foi administrada em quatro frações de 100cGy por local, a cada quatro dias e a uma profundidade de 0,2cm em cada fração. A betaterapia foi bem tolerada por todos os animais, sem ocorrência de catarata induzida pela radiação. Em todos os casos, houve um aumento dos sinais de inflamação da conjuntiva ao redor da neoformação, as quais cederam com o uso de anti-inflamatório tópico. Em dois cães houve a necessidade de tratamento cirúrgico, pois houve recorrência local do tumor aos 150 dias e aos 352 dias. Nos outros sete casos, o intervalo livre de doença variou entre 1.239 dias e 2.555 dias. Betaterapia usando 90Sr pode ser uma alternativa válida para o tratamento do CEC da terceira pálpebra em cães.

Comparison of conventional PCR, quantitative PCR, bacteriological culture and the Warthin Starry technique to detect Leptospira spp. in kidney and liver samples from naturally infected sheep from Brazil.

Leptospirosis is an infectious disease of worldwide importance. The development of diagnostic techniques allows sick animals to be identified, reservoirs to be eliminated and the disease prevented and controlled. The present study aimed to compare different techniques for diagnosing leptospirosis in sheep. Samples of kidney, liver and blood were collected from 465 animals that originated from a slaughterhouse. The sera were analyzed by the Microscopic Agglutination Test (MAT), and kidney and liver samples of seropositive animals were analyzed using four techniques: bacteriological culture, the Warthin Starry (WS) technique, conventional PCR (cPCR), and quantitative PCR (qPCR). With the MAT, 21 animals were positive (4.5%) to serovars Hardjo (n=12), Hebdomadis (n=5), Sentot (n=2), Wolfii (n=1) and Shermani (n=1). Titers were 100 (n=10), 200 (n=2), 400 (n=6) and 1600 (n=3). No animal was positive by bacteriological culture; four animals were positive by the WS technique in kidney samples; six animals were positive by cPCR in kidney samples; and 11 animals were positive by qPCR, eight of which in kidney samples and three in liver. The bacterial quantification revealed a median of 4.3 bacteria/µL in liver samples and 36.6 bacteria/µL in kidney samples. qPCR presented the highest sensitivity among the techniques, followed by cPCR, the WS technique and bacteriological culture. These results indicate that sheep can carry leptospires of the Sejroe serogroup, and demonstrate the efficiency of quantitative PCR to detect Leptospira spp. in tissue samples.

Occurrence and expression of p53 suppressor gene and c-Myc oncogene in dog eyelid tumors.

PURPOSE: To detect the occurrence and expression of the suppressor gene p53 and of the oncogene c-Myc in eyelid tumors of dogs using the PCR, RT-PCR, PCR-ELISA and RT-PCR-ELISA techniques. These genes have not been described in dog eyelid tumors before. METHODS: Nine samples of eyelid or third eyelid epithelial tumors were obtained from the archives of the Department of Veterinary Pathology. Tumor diagnosis was confirmed by evaluation of hematoxylin-eosin stained sections, and immunohistochemistry for cytokeratin AE1/AE3 and vimentin V9. A canine mammary tumor was used for positive control. Agarose gel electrophoresis, PCR-ELISA and RT-PCR-ELISA were used to detect p53 and c-Myc genes. RESULTS: The occurrence of p53 was detected in most of the eyelid tumors and third eyelid tumors studied (88.8%, n = 8) and was expressed in 75% of the positive samples, as indicated by ELISA. The c-Myc gene was found in 77.7% (n = 7) of the samples and was expressed in eight samples. CONCLUSIONS: Eyelid and third eyelid tumors of dogs express both the p53 and the c-Myc genes as shown by PCR and RT-PCR. However, PCR ELISA and RT-PCR ELISA were more efficient in assessing occurrence and expression of these genes because they identified amplified products that were not detected by agarose gel electrophoresis.

Detection of Toxoplasma gondii oocysts in environmental samples from public schools.

The number of Toxoplasma gondii oocysts that can be found in random environmental samples is probably low; in addition, these cysts may be confused with Hammondia spp. and Neospora spp. oocysts. The aim of the present work was to evaluate the presence of T. gondii oocysts in the soil of public elementary schools in the northwest area of the state of São Paulo, Brazil using mouse bioassays. A comparison was made between the different available bioassay techniques, such as squash, histopathology, immunohistochemistry and indirect fluorescent antibody test (IFAT). T. gondii was isolated by bioassay in mice (squash brain samples) from 22.58% (7/31) of the school playgrounds. Immunohistochemistry and IFAT showed positive results in 32.26% (10/31) and 25.80% (8/31) of samples, respectively. The sensitivity and specificity of the immunohistochemistry method were 85.71% and 83.33%, respectively. The IFAT results showed 100% sensitivity and 95.83% specificity. The presence of T. gondii was not detected in histopathological examinations. The results of the present study strongly suggest that T. gondii oocysts are widely distributed in elementary public schools in the region that was evaluated, likely constituting the main contamination source for these children. Educational programs directed at reducing environmental contamination with T. gondii would eventually lower the cost of treating humans for clinical toxoplasmosis. It is also possible to conclude that the use of IFAT in mouse bioassays can be recommended without the need for brain cysts research, which is extremely difficult and laborious.

A novel in situ polymerase chain reaction hybridisation assay for the direct detection of bovine herpesvirus type 5 in formalin-fixed, paraffin-embedded tissues.

An in situ polymerase chain reaction (IS-PCR) hybridisation assay was carried out on the brains of 20 cattle infected naturally with bovine herpesvirus type 5 (BoHV-5). Sections from the olfactory bulb and the frontal cortex of each sample were analysed using IS-PCR followed by hybridisation targeting the BoHV-5 US9 gene using a biotinylated primer. Each of the IS-PCR and hybridisation steps was optimised, and three different methods for detecting the virus were used. No false positive signals were observed in any negative control sample (n=20), resulting in a specificity of 100%. The results of IS-PCR hybridisation analysis of the olfactory bulb and the frontal cortex be compared directly with the results obtained using virus isolation, and the specificity and sensitivity were calculated. The most suitable method of visualisation was the peroxidase/3'-3-diaminobenzidine (DAB) detection system coupled with the use of the fluorescent dye Cy3. Using either of these methods, 80% of the positive samples (16 out of 20 samples) were identified using olfactory bulb sections. This is the first report using IS-PCR hybridisation for the direct detection of BoHV-5 DNA in clinical samples, and it provides an additional method for veterinary virology.

Experiencia clínica del uso de la tepoxalina en el tratamiento conservativo de la ruptura del ligamento cruzado craneal en perros / Clinical trial of tepoxalin use in cranial cruciate ligament ruptura conservative treatment in dogs

La ruptura de ligamento cruzado craneal (RLCC) se caracteriza por la laceración parcial o total del ligamento cruzado craneal. Pacientes con laceraciones agudas del ligamento (parcial o total) presentan claudicación aguda con incapacidad total o parcial de sustentación del peso en el miembro afectado. La terapia clínica con principios conservadores debe ser empleada previamente al tratamiento quirúrgico. La tepoxalina es un antiinflamatorio no esteroidal, indicado en la reducción de procesos inflamatorios y alivio del dolor causado por perturbaciones musculo-esqueléticas agudas y crónicas. Este medicamento promueve el bloqueo balanceado de la ruptura del ácido araquidónico por las vías de la lipoxigenasa y cicloxigenasa de la cascada de la inflamación. El presente trabajo tiene por objetivo evaluar la eficacia clínica del uso de la tepoxalina en el tratamiento conservativo de la RLLC, por medio de test clínicos específicos, examen radiográfico y análisis citológico del líquido articular. Diez perros con RLCC, macos y hembras, fueron incluidos en este estudio y recibieron dosis diarias de 10mg/kg de tepoxalina durante veintiocho días. La evaluación de la claudicación, los test de arco de movimiento en extensión máxima y digito-presión, así como el análisis del liquido articular después del periodo de tratamiento, permitieron concluir la efeicacia de la tepoxalina en el tratamiento conservativo de la RLCC. Su uso posibilitó el retorno a la deambulación normal en todos los casos tratados, siendo considerada una opción más de AINE para el tratamiento conservativo de esta artropatía...

Natural latex graft in lamellar and penetrating sclerectomies in rabbits / Enxerto de látex natural na cicatrização de esclerectomias lamelar e penetrante em coelhos

The objective of study was to investigate the effects of natural latex with 0.1 percent of polylysine on lamellar and penetrating scleroctomies in rabbits. Two groups of twelve rabbits each (lamellar GI and penetrating GII) were studied. Scleral square incisions near the limbus were performed on the left eye of each animal. The latex biomembrane was fixed to the recipient sclera and it covered with a conjunctival flap. The clinical evaluations were followed for 60 days. Aplannation tonometry, binocular indirect ophthalmoscopy and slit-lamp biomicroscopy were performed during evaluation. Bright field microscopy and polarization microscopy were employed. Blepharospasm, graft infection, mucoid ocular discharge and chemosis were not observed in either treatment group. The conjunctival hyperemia varied from moderate to hardly noticeable. The postoperative IOP was not statiscally significant, comparing to the preoperative IOP, for GI and GII. The histopathology by polarization microscopy showed that the neoformed tissue was primarily dependent on adjacent vascularized tissues and was constituted by collagen type III. Both groups presented optimum graft adhesion to the receiving sclera. The natural latex biomembrane with 0.1 percent polylysine constitutes a new alternative for scleral reconstruction. Furthermore, this is a durable material, easy to obtain and manipulate.

Este trabalho teve como objetivo investigar os efeitos do látex natural com polilisina a 0,1 por cento na cicatrização de esclerectomias lamelar e penetrante em coelhos. Foram estudados dois grupos de 12 coelhos (GI - lamelar e GII - penetrante). As esclerectomias foram realizadas no olho esquerdo de cada animal. A biomembrana de látex foi fixada à esclera receptora e foi recoberta com conjuntiva bulbar. As avaliações clínicas foram realizadas durante 60 dias. Para tal, empregaram-se a tonometria de aplanação, a oftalmoscopia indireta binocular e a biomicroscopia em lâmpada de fenda. Realizou-se análise histopatológica das escleras em microscopia de luz e sob luz polarizada. Não houve blefarospasmo, secreção ocular mucóide, quemose ou sinais de infecção do enxerto em ambos os grupos. A hiperemia conjuntival variou de moderada a tênue. Não houve diferença estatisticamente significativa entre a pressão intraocular do período pós-operatório e do período pré-operatório, para o GI e o GII. A histopatologia por microscopia de polarização demonstrou que o tecido neoformado foi oriundo de tecidos vascularizados adjacentes e constituídos por colágeno do tipo III. Os dois grupos apresentaram ótima adesão do enxerto de látex à esclera receptora. A biomembrana de látex natural com polilisina a 0,1 por cento representa uma alternativa para a reconstrução escleral, além de apresentar fácil obtenção, manuseio e durabilidade.

Comparative evaluation of enzyme-linked immunosorbent assay based on crude and purified antigen in the diagnosis of canine visceral leishmaniasis in symptomatic and oligosymptomatic dogs.

This study evaluated the performance of crude total antigen (CTA) and fucose-mannose ligand antigen (FML) in an enzyme-linked immunosorbent assay for diagnosis of canine visceral leishmaniasis (CVL). The assays used sera from known negative controls (n=30), clinically symptomatic (n=30) and oligosymptomatic (n=30) parasitologically proven infection (by microscopy). Aspirates of popliteal lymph node from infected canines were colleted to score parasitism and compared with the ELISA results. The study indicated that FML used in ELISA provided high sensitivity for detecting oligosymptomatic dogs (90%) and CTA showed greater sensitivity than FML for symptomatic canines (90%). In oligosymptomatic dogs, specificity was 100% for CTA-ELISA, but in symptomatic dogs, FML specificity was higher (96.7%) than CTA-ELISA (93.3%). A significant correlation was observed between the degree of parasitism and the results obtained in CTA-ELISA. Since no available antigen offers 100% specificity and sensitivity for CVL diagnosis, the choice of antigen used must depend on the aim of the investigation.

Direct detection of infectious bursal disease virus from clinical samples by in situ reverse transcriptase-linked polymerase chain reaction.

The presence of the very virulent (vv) Brazilian strain of infectious bursal disease virus (IBDV) was determined in the bursa of Fabricius, thymus and liver of 2-week-old broilers from a flock with a higher than expected mortality. For this purpose, a direct in situ reverse transcriptase (RT)-linked polymerase chain reaction (PCR) method was developed using specific primers for vvIBDV. Unlabelled forward and reverse biotinylated oligonucleotides were used for RT-PCR in a one-step method and the respective products were revealed by a direct enzymatic reaction. The results were compared with those obtained by standard RT-PCR using general primers for IBDV and virus isolation. The virus isolation, RT-PCR and in situ RT-PCR revealed positive results on the bursa of Fabricius in 86%, 80% and 100%, respectively. The in situ RT-PCR detected vvIBDV in all tested thymus and liver samples, whereas the standard RT-PCR detected virus in 80% and 90% of the samples, respectively. After three consecutive passages on chicken embryonated eggs, IBDV was isolated from 64% of the thymus samples and 30% of the liver samples. In the present study, no classical or antigenic variants of IBDV were detected. The developed in situ RT-PCR assay was able to detect the very virulent strain of IBDV with a higher sensitivity than the conventional RT-PCR and virus isolation.

Oral cysticercosis: case report.

Frequent in developing countries, cysticercosis is a parasitic infection that rarely involves the mouth. This study reports a case of oral cysticercosis in a 13-year-old female patient who had an asymptomatic nodule in the right labial mucosa. An excisional biopsy was carried out and the histopathologic examination revealed a cystic space containing a Taenia solium larva.

Detection of turkey coronavirus in commercial turkey poults in Brazil.

Poult enteritis complex has been incriminated as a major cause of loss among turkey poults in other countries. We have observed this in Brazil, associated with diarrhoea, loss of weight gain and, commonly, high mortality. In this study, we have used the reverse transcriptase polymerase chain reaction (RT-PCR) to detect turkey coronavirus (TCoV) in sick poults 30 to 120 days of age from a particular producer region in Brazil. The RT-PCR was applied to extracts of intestine tissue suspensions, and the respective intestinal contents, bursa of Fabrícius, faecal droppings and cloacal swabs. Primers were used to amplify the conserved 3' untranslated region of the genome, and the nucleocapsid protein gene of TCoV. Histopathological and direct immunohistochemical examinations were performed to detect TCoV antigen in infected intestine and bursa slides. All the results from stained tissues revealed lesions as described previously for TCoV infection. The direct immunohistochemical positive signal was present in all intestine slides. However, all bursa of Fabrícius tissues analysed were negative. RT-PCR findings were positive for TCoV in all faecal droppings samples, and in 27% of cloacal swabs. Finally, the best field material for TCoV diagnosis was faecal droppings and/or intestine suspensions.

Fungal pyogranulomatous encephalitis in a dog with leishmaniosis

É relatado um caso de encefalite piogranulomatosa em um cão fêmea de um ano de idade, da raça Fila Brasileiro. Ao exame macroscópico do cérebro, evidenciou-se área amolecida e hemorrágica no córtex frontal direito e na superfície de corte do hemisfério esquerdo, afetando a substância branca e áreas corticais profundas. O diagnóstico de encefalite piogranulomatosa micótica multifocal foi realizado através de exame histopatológico, que mostrou a presença de macrófagos, células gigantes, focos de hemorragia e hifas septadas de coloração marrom, com distribuição difusa e invadindo a luz de vasos. A identificação de formas amastigotas ôno imprintõ de linfonodo poplíteo confirmou o diagnóstico de leishmaniose. A infecção micótica no cérebro deste cão foi relacionada com a ocorrência concomitante de leishmaniose, uma doença imunossupressora.

Effect of age and abomasal puncture on peritoneal fluid, hematology, and serum biochemical analyses in young calves.

The goals of this study were to evaluate techniques for collection of peritoneal fluid from calves, establish reference ranges for fibrinogen in peritoneal fluid during the 1st month of life, and determine if abomasal puncture would alter peritoneal fluid or hematologic variables. Twenty-two healthy Holstein calves underwent 3 peritoneal fluid collections on day 1, day 15, and day 30 of age. Fibrinogen concentration in peritoneal fluid was 0.20 g/dL and 0.10 g/dL (P < .05) for day 1 and day 30, respectively, and 0.10 at day 15 (P > .05) for calves without abomasal puncture. Plasma fibrinogen concentration was 0.60 g/dL and 0.70 g/ dL (P < .05) for days 15 and 30, respectively, in calves without abomasal puncture. There were no significant differences (P < or = .05) in peritoneal fluid and peripheral blood total protein and fibrinogen concentrations, specific gravity, total and differential cell count, or erythrocyte counts between calves with or without abomasal puncture. We concluded that the reference ranges established for fibrinogen and total protein concentration are important for accurate evaluation of peritoneal fluid in calves for further comparison with similar-aged animals with gastrointestinal-tract or abdominal-cavity disease. Additionally, accidental abomasal puncture does not alter values of fibrinogen, total protein, and nucleated cell count in peritoneal fluid and does not cause apparent clinical abnormalities.