PESQUISA VETERINÁRIA BRASILEIRA

Abstract in English:

ABSTRACT.- David N., Hubner S.O., Riet-Correa F., Halfen D. & Lemos R.A. 2007. Reactivation of latent bovine herpesvirus type 5 in cattle with polioencephalomalacia induced by ammonium sulphate. Pesquisa Veterinária Brasileira 27(10):435-441. Hospital Veterinário, CSTR, Universidade Federal de Campina Grande, Campus de Patos, 58700-000 Patos, PB, Brazil. E-mail: franklin.riet@pesquisador.cnpq.br In the state Mato Grosso do Sul, Brazil, outbreaks of meningoencephalitis by BoHV-5 and polioencephalomalacia (PEM) display similar epidemiological features, suggesting that meningoencephalitis may be associated with reactivation of a latent BoHV-5 infection, during the development of PEM. To test this hypothesis, four 7-8 months old steers negative for BoHV-5 antibodies were inoculated intranasally with BoHV-5 and received amprolium from day 35 to day 105 after inoculation. Because PEM was not produced during this period, ammonium sulphate was given from day 114 to day 180 after inoculation. Two uninfected control steers received amprolium and ammonium sulphate for the same periods. All inoculated cattle developed antibodies against BoHV-5 after inoculation and the virus was isolated from nasal swabs, indicating that they were infected. Two inoculated steers had clinical signs of PEM after 118 and 146 days after virus inoculation. One was euthanized after a clinical manifestation period of seven days and had severe lesions of PEM and meningoencephalitis. BoHV-5 was isolated from the central nervous system of this animal. The other animal recovered but continued to manifest chronic signs of PEM and was euthanatized. On histological examination, the cerebral cortex, caudate nucleus and thalamus had multifocal areas of malacia and mild meningoencephalitis of the cortex. BoHV-5 was not isolated from the brain. One uninfected control steer had signs of neurological disease on day 158 and had lesions of PEM without meningoencephalitis at necropsy. The simultaneous production of PEM and diffuse meningoencephalitis, with isolation of BoHV-5, in one steer treated with ammonium sulphate, 118 days after BoHV-5 inoculation, suggests that latent BoHV-5 was reactivated in this animal submitted to experimental induction of PEM.

• Polioencephalomalacia bovine herpesvirus type 5 viral reactivation cattle

Abstract in Portuguese:

ABSTRACT.- David N., Hubner S.O., Riet-Correa F., Halfen D. & Lemos R.A. 2007. Reactivation of latent bovine herpesvirus type 5 in cattle with polioencephalomalacia induced by ammonium sulphate. Pesquisa Veterinária Brasileira 27(10):435-441. Hospital Veterinário, CSTR, Universidade Federal de Campina Grande, Campus de Patos, 58700-000 Patos, PB, Brazil. E-mail: franklin.riet@pesquisador.cnpq.br In the state Mato Grosso do Sul, Brazil, outbreaks of meningoencephalitis by BoHV-5 and polioencephalomalacia (PEM) display similar epidemiological features, suggesting that meningoencephalitis may be associated with reactivation of a latent BoHV-5 infection, during the development of PEM. To test this hypothesis, four 7-8 months old steers negative for BoHV-5 antibodies were inoculated intranasally with BoHV-5 and received amprolium from day 35 to day 105 after inoculation. Because PEM was not produced during this period, ammonium sulphate was given from day 114 to day 180 after inoculation. Two uninfected control steers received amprolium and ammonium sulphate for the same periods. All inoculated cattle developed antibodies against BoHV-5 after inoculation and the virus was isolated from nasal swabs, indicating that they were infected. Two inoculated steers had clinical signs of PEM after 118 and 146 days after virus inoculation. One was euthanized after a clinical manifestation period of seven days and had severe lesions of PEM and meningoencephalitis. BoHV-5 was isolated from the central nervous system of this animal. The other animal recovered but continued to manifest chronic signs of PEM and was euthanatized. On histological examination, the cerebral cortex, caudate nucleus and thalamus had multifocal areas of malacia and mild meningoencephalitis of the cortex. BoHV-5 was not isolated from the brain. One uninfected control steer had signs of neurological disease on day 158 and had lesions of PEM without meningoencephalitis at necropsy. The simultaneous production of PEM and diffuse meningoencephalitis, with isolation of BoHV-5, in one steer treated with ammonium sulphate, 118 days after BoHV-5 inoculation, suggests that latent BoHV-5 was reactivated in this animal submitted to experimental induction of PEM.

Abstract in English:

ABSTRACT.- Inoe A.P., Pereira F.C., Stopiglia A.J. & Da-Silva C.F. 2007. Pharmacological immunomodulation enhances peripheral nerve regeneration. Pesquisa Veterinária Brasileira 27(9):363-369. Departamento de Anatomia, Instituto de Ciências Biomédicas, Universidade de São Paulo, Cidade Universitária, Av. Prof. Lineu Prestes 2415, São Paulo, SP 055508-900, Brazil. E-mail: ana.paula@unipar.br To assess the effect of N-Acetylmuramyl-L-Alanyl-D-Isoglutamine MDP topically administrated on the regenerating peripheral neurons, twelve male C57BL/6J adult mice were equally distributed into three groups. Four mice underwent unilateral sciatic nerve transection and polyethylene tubulization, with a 4mm gap between the proximal and distal nerve stumps and were implanted with collagen + PBS (COL). Other four animals underwent the same surgical procedure but received collagen + MDP (COL/MDP) inside the prosthesis. Four animals were not operated and served as control group (NOR). After 4 weeks, the regenerated nerve cables were processed for total myelinated axon counting and myelinated fiber diameter measurement. The L5 dorsal root ganglion (DRG) was also removed and sectioned for sensory neurons counting and measurement. The results revealed significant difference (p<0.05) in axonal counting among the groups NOR (4,355±32), COL (1,869±289) and COL/MDP (2,430±223). There was a significant reduction in the axonal diameter in the operated groups (COL=3.38mm±1.16 and COL/MDP=3.54mm±1.16) compared to NOR (6.19mm±2.45). No difference was found in the number of DRG neurons between the experimental groups (COL=564±51; COL/MDP=514±56), which presented fewer sensory neurons compared to NOR (1,097±142). Data obtained indicate that locally applied MDP stimulates peripheral nerve regeneration in mice.

• N-Acetylmuramyl-L-Alanyl-D-Isoglutamine MDP nerve regeneration drug effect sciatic nerve surgery

Abstract in Portuguese:

ABSTRACT.- Inoe A.P., Pereira F.C., Stopiglia A.J. & Da-Silva C.F. 2007. Pharmacological immunomodulation enhances peripheral nerve regeneration. Pesquisa Veterinária Brasileira 27(9):363-369. Departamento de Anatomia, Instituto de Ciências Biomédicas, Universidade de São Paulo, Cidade Universitária, Av. Prof. Lineu Prestes 2415, São Paulo, SP 055508-900, Brazil. E-mail: ana.paula@unipar.br To assess the effect of N-Acetylmuramyl-L-Alanyl-D-Isoglutamine MDP topically administrated on the regenerating peripheral neurons, twelve male C57BL/6J adult mice were equally distributed into three groups. Four mice underwent unilateral sciatic nerve transection and polyethylene tubulization, with a 4mm gap between the proximal and distal nerve stumps and were implanted with collagen + PBS (COL). Other four animals underwent the same surgical procedure but received collagen + MDP (COL/MDP) inside the prosthesis. Four animals were not operated and served as control group (NOR). After 4 weeks, the regenerated nerve cables were processed for total myelinated axon counting and myelinated fiber diameter measurement. The L5 dorsal root ganglion (DRG) was also removed and sectioned for sensory neurons counting and measurement. The results revealed significant difference (p<0.05) in axonal counting among the groups NOR (4,355±32), COL (1,869±289) and COL/MDP (2,430±223). There was a significant reduction in the axonal diameter in the operated groups (COL=3.38mm±1.16 and COL/MDP=3.54mm±1.16) compared to NOR (6.19mm±2.45). No difference was found in the number of DRG neurons between the experimental groups (COL=564±51; COL/MDP=514±56), which presented fewer sensory neurons compared to NOR (1,097±142). Data obtained indicate that locally applied MDP stimulates peripheral nerve regeneration in mice.

Abstract in English:

ABSTRACT.- Claus M.P., Vivian D., Lunardi M., Alfieri A.F. & Alfieri A.A. 2007. [Phylogenetic analysis of bovine papillomavirus associated with skin warts in cattle herds from the state of Paraná.] Análise filogenética do papilomavírus bovino associado a lesões cutâneas em rebanhos do Estado do Paraná. Pesquisa Veterinária Brasileira 27(7):314-318. Laboratório de Virologia Animal, Departamento de Medicina Veterinária Preventiva, Centro de Ciências Agrárias, Universidade Estadual de Londrina, Cx. Postal 6001, Campus Universitário, Londrina, PR 86051-990, Brazil. E-mail: alfieri@uel.br Bovine papillomavirus (BPV) infection causes hyperplastic lesions in the cutaneous epithelium of cattle. Six types of BPV were classified in two sub-groups, being correlated to the anatomical regions of the infection and morphologic characteristics of the lesions. The present study was carried out to identify the types of BPV present in skin warts of cattle from the state of Paraná, Brazil. The generic primers FAP59 and FAP64 were used for amplification of a 478 bp fragment of BPV L1 gene in nine cutaneous papilloma samples obtained from six animals in four herds. In all papillomas examined, a product with the expected molecular size was amplified. Phylogenetic analysis of the PCR products identified BPV-2 in three samples, BPV-1 in one, and BPV-6 in five papillomas. BPV-6 was detected in cutaneous papillomas of the teat and in other body parts as well. In one animal, from which more than one sample was collected, a concomitant infection by BPV-1 and BPV-2 was identified. The five positive BPV-6 samples showed a nucleotide identity of 100% with the sequence of the reference strain available in GenBank. However, differences among BPV-2 and BPV-1 Brazilian samples and the respective reference sequences deposited in GenBank were observed. Molecular comparison of the two BPV-2 strains identified showed the involvement of two viral variants. This study revealed the diversity of BPV types circulating in the state of Paraná.

• Bovine papillomavirus cutaneous papillomatosis phylogeny polymerase chain reaction

Abstract in Portuguese:

ABSTRACT.- Claus M.P., Vivian D., Lunardi M., Alfieri A.F. & Alfieri A.A. 2007. [Phylogenetic analysis of bovine papillomavirus associated with skin warts in cattle herds from the state of Paraná.] Análise filogenética do papilomavírus bovino associado a lesões cutâneas em rebanhos do Estado do Paraná. Pesquisa Veterinária Brasileira 27(7):314-318. Laboratório de Virologia Animal, Departamento de Medicina Veterinária Preventiva, Centro de Ciências Agrárias, Universidade Estadual de Londrina, Cx. Postal 6001, Campus Universitário, Londrina, PR 86051-990, Brazil. E-mail: alfieri@uel.br Bovine papillomavirus (BPV) infection causes hyperplastic lesions in the cutaneous epithelium of cattle. Six types of BPV were classified in two sub-groups, being correlated to the anatomical regions of the infection and morphologic characteristics of the lesions. The present study was carried out to identify the types of BPV present in skin warts of cattle from the state of Paraná, Brazil. The generic primers FAP59 and FAP64 were used for amplification of a 478 bp fragment of BPV L1 gene in nine cutaneous papilloma samples obtained from six animals in four herds. In all papillomas examined, a product with the expected molecular size was amplified. Phylogenetic analysis of the PCR products identified BPV-2 in three samples, BPV-1 in one, and BPV-6 in five papillomas. BPV-6 was detected in cutaneous papillomas of the teat and in other body parts as well. In one animal, from which more than one sample was collected, a concomitant infection by BPV-1 and BPV-2 was identified. The five positive BPV-6 samples showed a nucleotide identity of 100% with the sequence of the reference strain available in GenBank. However, differences among BPV-2 and BPV-1 Brazilian samples and the respective reference sequences deposited in GenBank were observed. Molecular comparison of the two BPV-2 strains identified showed the involvement of two viral variants. This study revealed the diversity of BPV types circulating in the state of Paraná.

Abstract in English:

ABSTRACT.- Rissi D.R., Rech R.R., Flores E.F., Kommers G.D. & Barros C.S.L. 2007. [Meningoencephalitis by bovine herpesvirus-5.] Meningoencefalite por herpesvírus bovino-5. Pesquisa Veterinária Brasileira 27(7):251-260. Departamento de Patologia, Universidade Federal de Santa Maria, 97105-900, Santa Maria, RS, Brazil. E-mail: claudioslbarros@uol.com.br Meningoencephalitis caused by bovine herpesvirus-5 (BoHV-5) is an often fatal, acute or subacute infectious disease that affects mainly young cattle under stressing conditions. The disease has been recognized in several Brazilian regions and in other parts of the world. BoHV-5 is a double stranded DNA virus member of the Herpesviridae family and subfamily Alphaherpesvirinae. The virus is characterized by rapid and lytic replication in cell cultures and by the ability to establish lifelong latent infection in sensory nerve ganglia of the host. BoHV-5 is transmitted mainly by direct and indirect contact and replicates acutely in the oral, nasal, oropharingeal or ocular mucosae. After primary replication, the virus invades nerve endings and is transported to the neuron cell bodies of the sensory ganglia where it replicates actively and/or establishes latency. Viral invasion of the brain may result in massive virus replication and production of neurological disease. Virtually all cattle developing neurological disease die of meningoencephalitis; yet the infection may be subclinical in some animals. These animals recover and become latently infected. Viral dissemination within a herd is facilitated by conditions such as crowding, introduction of cattle from other herds and weaning of calves in ages that coincide with decrease of passive immunity. Certain natural or induced conditions may reactivate the latent virus and favor its transmission and dissemination to other susceptible individuals. The disease may occur as outbreaks or as sporadic cases, with morbidity rates ranging of 0.05%-5%; lethality is almost always 100%. Clinical signs include depression, nasal and ocular discharge, grinding of teeth, circling, blindness, fever, paddling movements, disphagia, abdominal pain, nystagmus, muscle tremors, drooling, incoordinated gait, opisthotonus, head pressing, falls and convulsions. Clinical course is usually 1-15 days. Necropsy findings may be absent but often there is swollen of the rostral portions of the cerebral cortex and flattening of gyri, with softening and segmental yellow discoloration (malacia). As the disease progresses the affected areas become gelatinous and grey and, in advanced cases, there is segmental loss of the cerebral cortex of the frontal lobe of the brain (residual lesion). In several cases there is malacia of the basal nuclei and of the thalamus. Histologically, there is necrotizing non-suppurative meningoencephalitis affecting mainly the cerebral cortex of the frontal lobe associated with eosinophilic intranuclear inclusion bodies in neurons and astrocytes, although the frequency of the inclusion bodies is inconsistent. The diagnosis of meningoencephalitis by BoHV-5 should be based on epidemiology, clinical signs, necropsy and histological findings. The diagnosis should be confirmed by viral isolation in cell culture and/or by detection of viral antigens in brain sections or in exfoliated cells from nasal secretions. The identification and characterization of BoHV-5 can be done by the use of monoclonal antibodies, polymerase chain reaction (PCR) and/or by restriction enzyme analysis of the viral genome. There is no specific treatment for the disease. As BoHV-1 and BoHV-5 are antigenically related, vaccination using BoHV-1 vaccines may be recommended as a means of reducing the losses caused by BoHV-5 infection, mainly during outbreaks of neurologic disease. Additionally, measures such as serologic testing of new additions to the herd; and management practices to prevent stress and to reduce conditions for virus dissemination among animals may help in reducing the incidence and the consequences of BoHV-5 infection and disease.

• Bovine herpesvirus-5 BoHV-5 BHV-5 meningoencephalitis viral diseases diseases of cattle neuropathology

Abstract in Portuguese:

ABSTRACT.- Rissi D.R., Rech R.R., Flores E.F., Kommers G.D. & Barros C.S.L. 2007. [Meningoencephalitis by bovine herpesvirus-5.] Meningoencefalite por herpesvírus bovino-5. Pesquisa Veterinária Brasileira 27(7):251-260. Departamento de Patologia, Universidade Federal de Santa Maria, 97105-900, Santa Maria, RS, Brazil. E-mail: claudioslbarros@uol.com.br Meningoencephalitis caused by bovine herpesvirus-5 (BoHV-5) is an often fatal, acute or subacute infectious disease that affects mainly young cattle under stressing conditions. The disease has been recognized in several Brazilian regions and in other parts of the world. BoHV-5 is a double stranded DNA virus member of the Herpesviridae family and subfamily Alphaherpesvirinae. The virus is characterized by rapid and lytic replication in cell cultures and by the ability to establish lifelong latent infection in sensory nerve ganglia of the host. BoHV-5 is transmitted mainly by direct and indirect contact and replicates acutely in the oral, nasal, oropharingeal or ocular mucosae. After primary replication, the virus invades nerve endings and is transported to the neuron cell bodies of the sensory ganglia where it replicates actively and/or establishes latency. Viral invasion of the brain may result in massive virus replication and production of neurological disease. Virtually all cattle developing neurological disease die of meningoencephalitis; yet the infection may be subclinical in some animals. These animals recover and become latently infected. Viral dissemination within a herd is facilitated by conditions such as crowding, introduction of cattle from other herds and weaning of calves in ages that coincide with decrease of passive immunity. Certain natural or induced conditions may reactivate the latent virus and favor its transmission and dissemination to other susceptible individuals. The disease may occur as outbreaks or as sporadic cases, with morbidity rates ranging of 0.05%-5%; lethality is almost always 100%. Clinical signs include depression, nasal and ocular discharge, grinding of teeth, circling, blindness, fever, paddling movements, disphagia, abdominal pain, nystagmus, muscle tremors, drooling, incoordinated gait, opisthotonus, head pressing, falls and convulsions. Clinical course is usually 1-15 days. Necropsy findings may be absent but often there is swollen of the rostral portions of the cerebral cortex and flattening of gyri, with softening and segmental yellow discoloration (malacia). As the disease progresses the affected areas become gelatinous and grey and, in advanced cases, there is segmental loss of the cerebral cortex of the frontal lobe of the brain (residual lesion). In several cases there is malacia of the basal nuclei and of the thalamus. Histologically, there is necrotizing non-suppurative meningoencephalitis affecting mainly the cerebral cortex of the frontal lobe associated with eosinophilic intranuclear inclusion bodies in neurons and astrocytes, although the frequency of the inclusion bodies is inconsistent. The diagnosis of meningoencephalitis by BoHV-5 should be based on epidemiology, clinical signs, necropsy and histological findings. The diagnosis should be confirmed by viral isolation in cell culture and/or by detection of viral antigens in brain sections or in exfoliated cells from nasal secretions. The identification and characterization of BoHV-5 can be done by the use of monoclonal antibodies, polymerase chain reaction (PCR) and/or by restriction enzyme analysis of the viral genome. There is no specific treatment for the disease. As BoHV-1 and BoHV-5 are antigenically related, vaccination using BoHV-1 vaccines may be recommended as a means of reducing the losses caused by BoHV-5 infection, mainly during outbreaks of neurologic disease. Additionally, measures such as serologic testing of new additions to the herd; and management practices to prevent stress and to reduce conditions for virus dissemination among animals may help in reducing the incidence and the consequences of BoHV-5 infection and disease.

Abstract in English:

ABSTRACT.- Sant’Ana F.J.F., Garcia E.C., Rabelo R.E., Costa Y.L., Schild A.L. & Riet-Correa F. 2007. [Oropharyngeal and neurologic lesions in sheep associated with the use of drenching guns.] Lesões orofaríngeas e neurológicas em ovinos associadas ao uso de pistolas dosificadoras. Pesquisa Veterinária Brasileira 27(7):282-286. Laboratório de Patologia Veterinária, Universidade Federal de Goiás, Campus Jataí, 75800-000 Jataí, GO, Brazil. E-mail: santanafjf@yahoo.com Six outbreaks of oropharyngeal lesions caused by drenching guns in sheep are reported. In 2 outbreaks in Rio Grande do Sul the morbidity was 6.25% and 15%, and the lesions observed after antihelmintic treatment in adult sheep were characterized by a fistula extending from mouth to skin of the face. All sheep died or were euthanatized after continuous weight loss for up to 8 months. Another outbreak in the same State affected 26 out of 360 adult sheep after being dosed with molasses, causing lesions in the oral mucosa followed by malignant edema of the oral and subcutaneous tissues. All sheep died within 10 days after treatment. Another case observed in Rio Grande do Sul affected one out of 88 adult ewes. The animal died with hemorrhages from the nose and mouth 8 days after antihelmintic treatment. At necropsy an ulcerated lesion covered by a clot of fibrine was found in the oral mucosa. The lesion had a fistulated tract with purulent exudate extending through the soft tissues in direction to the right eye. Two outbreaks occurred in the State of Goiás 16-20 days after antihelmintic treatment in 5-9 months old lambs. Morbidity was 11.43% and 15.62% and fatality rate was 7.14% and 9.38%. Main clinical signs were nervous signs caused by abscesses with a diameter of 0.5-3cm, containing purulent exudates in the oropharynx, soft palate and adjacent muscles extending to the meninges through the atlantoaxial joint. Not-purulent meningitis was observed histologically. The need to use proper drenching guns and pay attention during dosing to avoid lesions of the oropharynx is pointed out.

• Oropharyngeal abscess neurologic alterations drenching guns sheep

Abstract in Portuguese:

ABSTRACT.- Sant’Ana F.J.F., Garcia E.C., Rabelo R.E., Costa Y.L., Schild A.L. & Riet-Correa F. 2007. [Oropharyngeal and neurologic lesions in sheep associated with the use of drenching guns.] Lesões orofaríngeas e neurológicas em ovinos associadas ao uso de pistolas dosificadoras. Pesquisa Veterinária Brasileira 27(7):282-286. Laboratório de Patologia Veterinária, Universidade Federal de Goiás, Campus Jataí, 75800-000 Jataí, GO, Brazil. E-mail: santanafjf@yahoo.com Six outbreaks of oropharyngeal lesions caused by drenching guns in sheep are reported. In 2 outbreaks in Rio Grande do Sul the morbidity was 6.25% and 15%, and the lesions observed after antihelmintic treatment in adult sheep were characterized by a fistula extending from mouth to skin of the face. All sheep died or were euthanatized after continuous weight loss for up to 8 months. Another outbreak in the same State affected 26 out of 360 adult sheep after being dosed with molasses, causing lesions in the oral mucosa followed by malignant edema of the oral and subcutaneous tissues. All sheep died within 10 days after treatment. Another case observed in Rio Grande do Sul affected one out of 88 adult ewes. The animal died with hemorrhages from the nose and mouth 8 days after antihelmintic treatment. At necropsy an ulcerated lesion covered by a clot of fibrine was found in the oral mucosa. The lesion had a fistulated tract with purulent exudate extending through the soft tissues in direction to the right eye. Two outbreaks occurred in the State of Goiás 16-20 days after antihelmintic treatment in 5-9 months old lambs. Morbidity was 11.43% and 15.62% and fatality rate was 7.14% and 9.38%. Main clinical signs were nervous signs caused by abscesses with a diameter of 0.5-3cm, containing purulent exudates in the oropharynx, soft palate and adjacent muscles extending to the meninges through the atlantoaxial joint. Not-purulent meningitis was observed histologically. The need to use proper drenching guns and pay attention during dosing to avoid lesions of the oropharynx is pointed out.

Abstract in English:

ABSTRACT.- Cazerta S.M.M., Miglino M.A., Marques R.S., Vulcano M. & Pereira F.T.V. 2007. [Characterization of hemophagous areas of the bovine placenta.] Caracterização das áreas hemófagas da placenta bovina. Pesquisa Veterinária Brasileira 26(6):229-235. Faculdade de Zootecnia, Campus Dracena, Unesp, Rod. Com. João Ribeiro de Barros, SP294 Km 651, Dracena, SP 17900-000, Brazil. E-mail: fverechia@dracena.unesp.br The specific region of maternal fetal interface needs to be clarified and corresponds to the “arcade zone” of sheep and goat placentomes. In small ruminants that area is also characterized by macroscopic blood extravasation (hemophagous areas). This occurs possibly because the iron is transferred to the embryo by trophoblastic erytrophagocytosis in these hemophagous placental areas. In order to investigate the hemophagous placental areas in cattle, placentomes of 34 pregnant Bos indicus cows (2-3, 4-6, 7-8 and 9 months of gestation) were analyzed. They were fixed by perfusion with 10% formaldehyde aqueous solution and 4% paraformoldehyde in PBS, pH 7.4, 0.1M, processed and stained for light microscopy and histochemistry. The methodology used allowed to observe placental haematomes between the uterine and trophoblastic epithelium since 3 months of pregnancy. Erythrocytes had been found in the trophoblastic cells, elucidating the erytrophagocytosis. Through Perl’s histochemical reaction it was possible to prove the existence of ferric iron in the trophoblastic cells. The PAS reaction was positive staining mucoid substance in the epithelial cells, especially in the binucleate cells of the fetal epithelium. Based on histology and histochemistry, it can be inferred that the hemophagous areas are important sites for iron transfer in the bovine placenta.

• Erythrophagocytosis hemophagous areas bovine placenta reproduction trophoblast

Abstract in Portuguese:

ABSTRACT.- Cazerta S.M.M., Miglino M.A., Marques R.S., Vulcano M. & Pereira F.T.V. 2007. [Characterization of hemophagous areas of the bovine placenta.] Caracterização das áreas hemófagas da placenta bovina. Pesquisa Veterinária Brasileira 26(6):229-235. Faculdade de Zootecnia, Campus Dracena, Unesp, Rod. Com. João Ribeiro de Barros, SP294 Km 651, Dracena, SP 17900-000, Brazil. E-mail: fverechia@dracena.unesp.br The specific region of maternal fetal interface needs to be clarified and corresponds to the “arcade zone” of sheep and goat placentomes. In small ruminants that area is also characterized by macroscopic blood extravasation (hemophagous areas). This occurs possibly because the iron is transferred to the embryo by trophoblastic erytrophagocytosis in these hemophagous placental areas. In order to investigate the hemophagous placental areas in cattle, placentomes of 34 pregnant Bos indicus cows (2-3, 4-6, 7-8 and 9 months of gestation) were analyzed. They were fixed by perfusion with 10% formaldehyde aqueous solution and 4% paraformoldehyde in PBS, pH 7.4, 0.1M, processed and stained for light microscopy and histochemistry. The methodology used allowed to observe placental haematomes between the uterine and trophoblastic epithelium since 3 months of pregnancy. Erythrocytes had been found in the trophoblastic cells, elucidating the erytrophagocytosis. Through Perl’s histochemical reaction it was possible to prove the existence of ferric iron in the trophoblastic cells. The PAS reaction was positive staining mucoid substance in the epithelial cells, especially in the binucleate cells of the fetal epithelium. Based on histology and histochemistry, it can be inferred that the hemophagous areas are important sites for iron transfer in the bovine placenta.

Abstract in English:

ABSTRACT.- Agreste F.R., Bombonato P.P. & Hernandez-Blazquez, F.J. 2007. [Morphometric characteristics of thymus development in dogs.] Características morfométricas do desenvolvimento do timo em cães. Pesquisa Veterinária Brasileira 27(6):236-240. Departamento de Cirurgia, Setor de Anatomia dos Animais Domésticos e Silvestres, Universidade de São Paulo, Avenida Prof. Dr. Orlando Marques de Paiva 87, São Paulo, SP 05508-000, Brazil. E-mail: bombonat@usp.br The thymus is an organ with great immunological relevance during foetus life and neonatal period, It is the predecessor of lymphopoiesis and shows high lymphopoietic activity, being also the largest lymphoid organ during intrauterine life and at birth. In this study, the morphological aspects of the prenatal development of dog thymus were described in foetus of different ages and sexes. Twenty-four foetus of mongrel domestic dogs, males and females, were divided into four age groups. The thymus presented two lobes linked by a rose-colored tissue connection that was localized in the cranial mediastinal space. The cranial portion protrudes slightly beyond the first pair of ribs. The cells that formed the thymus parenchyma were well organized. Concentric aggregates named Hassal corpuscles were observed, involved by a thin capsule of connective tissue thickened by adipose tissue. The vessels size gradually increased with the foetus age, being larger in animals that were closer to birth. The volume (Vref) and size parameters of female thymus were larger than in males.

• Thymus vascularization lymphatic system morphometry dogs

Abstract in Portuguese:

ABSTRACT.- Agreste F.R., Bombonato P.P. & Hernandez-Blazquez, F.J. 2007. [Morphometric characteristics of thymus development in dogs.] Características morfométricas do desenvolvimento do timo em cães. Pesquisa Veterinária Brasileira 27(6):236-240. Departamento de Cirurgia, Setor de Anatomia dos Animais Domésticos e Silvestres, Universidade de São Paulo, Avenida Prof. Dr. Orlando Marques de Paiva 87, São Paulo, SP 05508-000, Brazil. E-mail: bombonat@usp.br The thymus is an organ with great immunological relevance during foetus life and neonatal period, It is the predecessor of lymphopoiesis and shows high lymphopoietic activity, being also the largest lymphoid organ during intrauterine life and at birth. In this study, the morphological aspects of the prenatal development of dog thymus were described in foetus of different ages and sexes. Twenty-four foetus of mongrel domestic dogs, males and females, were divided into four age groups. The thymus presented two lobes linked by a rose-colored tissue connection that was localized in the cranial mediastinal space. The cranial portion protrudes slightly beyond the first pair of ribs. The cells that formed the thymus parenchyma were well organized. Concentric aggregates named Hassal corpuscles were observed, involved by a thin capsule of connective tissue thickened by adipose tissue. The vessels size gradually increased with the foetus age, being larger in animals that were closer to birth. The volume (Vref) and size parameters of female thymus were larger than in males.

Abstract in English:

ABSTRACT.- Rodrigues M.A.M., Oliveira D.A., Taketomi E.A. & Hernandez-Blazquez F.J. 2007. IgA production, coliforms analysis and intestinal mucosa morphology of piglets that received probiotics with viable or inactivated cells. Pesquisa Veterinária Brasileira 27(6):241-245. Departamento de Cirurgia, Setor de Anatomia, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Av. Prof. Dr. Orlando Marques de Paiva 87, São Paulo, SP 05508-900, Brazil. Email: fjhblazq@usp.br Two types of probiotics were used in piglets. One product is a mixed culture of viable Lactobacillus acidophilus, Enterococcus faecium e Bifidobacterium bifidum. The second product is composed of inactivated Lactobacillus acidophilus cells. The piglets received two weekly oral doses for 30 days while a control group did not receive probiotics. All piglets were euthanized at the 30th day of life and the mesenteric lymph nodes, the small intestine, and blood samples were collected. The tissue samples were studied by light microscopy and the blood serum was analyzed by ELISA method. The treatment with the probiotic with viable cells produced higher serum levels of IgA (P<0.05) and more IgA expressing cells were found in the mesenteric lymph nodes than observed in the inactivated cells treatment or control groups (P<0.05). Also, intestinal villi were longer, crypts were deeper (P<0.05) and fecal coliform count was lower than found in the inactivated product (P<0.05). These results suggest that viable probiotics are more efficient than inactivated probiotics to induce immunostimulation and intestinal modifications in piglets, thus improving their health and development.

• Probiotics IgA piglets swine weaning Lactobacillus intestine

Abstract in Portuguese:

ABSTRACT.- Rodrigues M.A.M., Oliveira D.A., Taketomi E.A. & Hernandez-Blazquez F.J. 2007. IgA production, coliforms analysis and intestinal mucosa morphology of piglets that received probiotics with viable or inactivated cells. Pesquisa Veterinária Brasileira 27(6):241-245. Departamento de Cirurgia, Setor de Anatomia, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Av. Prof. Dr. Orlando Marques de Paiva 87, São Paulo, SP 05508-900, Brazil. Email: fjhblazq@usp.br Two types of probiotics were used in piglets. One product is a mixed culture of viable Lactobacillus acidophilus, Enterococcus faecium e Bifidobacterium bifidum. The second product is composed of inactivated Lactobacillus acidophilus cells. The piglets received two weekly oral doses for 30 days while a control group did not receive probiotics. All piglets were euthanized at the 30th day of life and the mesenteric lymph nodes, the small intestine, and blood samples were collected. The tissue samples were studied by light microscopy and the blood serum was analyzed by ELISA method. The treatment with the probiotic with viable cells produced higher serum levels of IgA (P<0.05) and more IgA expressing cells were found in the mesenteric lymph nodes than observed in the inactivated cells treatment or control groups (P<0.05). Also, intestinal villi were longer, crypts were deeper (P<0.05) and fecal coliform count was lower than found in the inactivated product (P<0.05). These results suggest that viable probiotics are more efficient than inactivated probiotics to induce immunostimulation and intestinal modifications in piglets, thus improving their health and development.

Abstract in English:

ABSTRACT.- Iturrizaga D.M., Verechia F.T., Santos T.C., Bombonato P.P., Teixeira D.G. & Miglino M.A. 2007. The materno-fetal interface in llama (Lama guanicoe glama). Pesquisa Veterinária Brasileira 27(6):221-228. Departamento de Cirurgia, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Av. Prof. Dr. Orlando Marques de Paiva 87, São Paulo, SP 05508-270, Brazil. E-mail: miglino@usp.br Samples from 9 llamas (28 through 36 weeks of gestation) were collected and fixed in 4% buffered paraformaldehyde (light microscopy) and in 2.5% buffered glutaraldehyde (transmission and scanning electron microscopy). The material was processed in paraplast and slides (5mm) were stained with HE, PAS, Masson-Trichrome, acid phosphatase and Perl´s. The uteroferrin was immunolocalized. The results show that llama placenta is chorioallantoic, diffuse, folded and epitheliochorial, and the fetus is covered with an epidermal membrane. The trophoblast cells have variable morphology: cubic, rounded and triangular cells, with cytoplasm containing PAS-positive granules. Binucleated cells with large cytoplasm and rounded nuclei, as well as giant trophoblastic cells with multiple nuclei were also observed. Numerous blood vessels were observed beneath the cells of the uterine epithelium and around the chorionic subdivided branches. Glandular activity was shown by PAS, Perl´s, and acid phosphatase positive reactions in the cytoplasm and glandular lumen, and by immunolocalization of the uteroferrin in the glandular epithelium. The uterine glands open in spaces formed by the areoles, which are filled by PAS-positive material. The llama fetus was covered by the epidermal membrane, composed of stratified epithelium, with up to seven layers of mono-, bi- or trinucleated cells. The high level of maternal and fetal vascularization surfaces indicates an intense exchange of substances across both surfaces. The metabolic activity shown in the uterine glands suggests an adaptation of the gestation to the high altitudes of the natural habitat of this species.

• Placenta llama trophoblast uteroferrin ephiteliochorial morphology

Abstract in Portuguese:

ABSTRACT.- Iturrizaga D.M., Verechia F.T., Santos T.C., Bombonato P.P., Teixeira D.G. & Miglino M.A. 2007. The materno-fetal interface in llama (Lama guanicoe glama). Pesquisa Veterinária Brasileira 27(6):221-228. Departamento de Cirurgia, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Av. Prof. Dr. Orlando Marques de Paiva 87, São Paulo, SP 05508-270, Brazil. E-mail: miglino@usp.br Samples from 9 llamas (28 through 36 weeks of gestation) were collected and fixed in 4% buffered paraformaldehyde (light microscopy) and in 2.5% buffered glutaraldehyde (transmission and scanning electron microscopy). The material was processed in paraplast and slides (5mm) were stained with HE, PAS, Masson-Trichrome, acid phosphatase and Perl´s. The uteroferrin was immunolocalized. The results show that llama placenta is chorioallantoic, diffuse, folded and epitheliochorial, and the fetus is covered with an epidermal membrane. The trophoblast cells have variable morphology: cubic, rounded and triangular cells, with cytoplasm containing PAS-positive granules. Binucleated cells with large cytoplasm and rounded nuclei, as well as giant trophoblastic cells with multiple nuclei were also observed. Numerous blood vessels were observed beneath the cells of the uterine epithelium and around the chorionic subdivided branches. Glandular activity was shown by PAS, Perl´s, and acid phosphatase positive reactions in the cytoplasm and glandular lumen, and by immunolocalization of the uteroferrin in the glandular epithelium. The uterine glands open in spaces formed by the areoles, which are filled by PAS-positive material. The llama fetus was covered by the epidermal membrane, composed of stratified epithelium, with up to seven layers of mono-, bi- or trinucleated cells. The high level of maternal and fetal vascularization surfaces indicates an intense exchange of substances across both surfaces. The metabolic activity shown in the uterine glands suggests an adaptation of the gestation to the high altitudes of the natural habitat of this species.

Abstract in English:

ABSTRACT.- Domingues-F. L.M. & Gioso M.A. 2007. [Comparison of the application of temporary endodontic dressing in teeth of dogs: histopathological and microbiological aspects.] Comparação entre sessão única e a utilização do “curativo de demora” no tratamento endodôntico em cães: aspectos histopatológicos e microbiológicos. Pesquisa Veterinária Brasileira 27(5):191-193. Departamento de Cirurgia da Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Av. Prof. Dr. Orlando Marques de Paiva 87, Bloco 8, Cidade Universitária, São Paulo, SP 05508-000, Brazil. E-mail: lesliedf@usp.br During a root canal therapy the microbiota is the major concern. Bacteria present in dentinal tubules, apical foramens and apical delta is related to failure of this procedure. Some studies reveal that during endodontic treatment in one session bacteria still remain in dental structures. The aim of the present study was to evaluate the endodontic treatment in one or two sessions, using calcium hydroxide plus camphorated paramonochlorophenol (CMCP) with temporary endodontic medicaments. Four dogs were used, and two animals were submitted the treatment in one session only and the others in two sessions. Premolar teeth were used in this study: second and third upper premolar teeth and second, third and fourth lower premolar teeth were divided into two groups. Opening of the pulp chamber was performed with burns, and the pulp was exposed for 60 days. After this period the group treated in only session was obtured with gutta-percha and zinc oxide and eugenol, and the group treated in two sessions received the CMCP that remained in the canal per 30 days and was afterwards obtured with the material of the other group. Clinical and radiographical control was performed twice a week, and after 60 days a small block containing the teeth, surrounding periapical tissues and the periodontium was removed for histological and microbiological study. Histological analysis revealed intense inflammatory response in both groups. Microbiological analysis showed a more severe microbial reduction in the group treated in one session. However, bacteria were still observed in the two groups, indicating that none of the treatments presented total efficiency.

• Apical delta endodontics gutta-percha calcium hydroxide dog camphorate paramonochlorophenol

Abstract in Portuguese:

ABSTRACT.- Domingues-F. L.M. & Gioso M.A. 2007. [Comparison of the application of temporary endodontic dressing in teeth of dogs: histopathological and microbiological aspects.] Comparação entre sessão única e a utilização do “curativo de demora” no tratamento endodôntico em cães: aspectos histopatológicos e microbiológicos. Pesquisa Veterinária Brasileira 27(5):191-193. Departamento de Cirurgia da Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Av. Prof. Dr. Orlando Marques de Paiva 87, Bloco 8, Cidade Universitária, São Paulo, SP 05508-000, Brazil. E-mail: lesliedf@usp.br During a root canal therapy the microbiota is the major concern. Bacteria present in dentinal tubules, apical foramens and apical delta is related to failure of this procedure. Some studies reveal that during endodontic treatment in one session bacteria still remain in dental structures. The aim of the present study was to evaluate the endodontic treatment in one or two sessions, using calcium hydroxide plus camphorated paramonochlorophenol (CMCP) with temporary endodontic medicaments. Four dogs were used, and two animals were submitted the treatment in one session only and the others in two sessions. Premolar teeth were used in this study: second and third upper premolar teeth and second, third and fourth lower premolar teeth were divided into two groups. Opening of the pulp chamber was performed with burns, and the pulp was exposed for 60 days. After this period the group treated in only session was obtured with gutta-percha and zinc oxide and eugenol, and the group treated in two sessions received the CMCP that remained in the canal per 30 days and was afterwards obtured with the material of the other group. Clinical and radiographical control was performed twice a week, and after 60 days a small block containing the teeth, surrounding periapical tissues and the periodontium was removed for histological and microbiological study. Histological analysis revealed intense inflammatory response in both groups. Microbiological analysis showed a more severe microbial reduction in the group treated in one session. However, bacteria were still observed in the two groups, indicating that none of the treatments presented total efficiency.