Effect of cryopreservation on sperm DNA fragmentation and apoptosis rates in the testicular tissue of domestic cats.

The objectives of the present study were to evaluate the damage caused by cryopreservation on sperm DNA and estimate the percentage of cell apoptosis in tissue after thawing. Testicles of cats were sectioned into of 0.3 cm3 and 0.5 cm3 fragments and evaluated for DNA damage using acridine orange and semi-quantitatively through histo-morphological and immunohistochemical methods (caspase-3). Other fragments were placed in cryotubes with diluent containing either 3% glycerol or 3% propanediol, and were cryopreserved. Evaluation using acridine orange indicated there was a difference with use of propanediol and glycerol on DNA damage in 0.5 cm3fragments, with the latter being more effective than the former for cryopreservation. Results from histomorphological evaluations indicated there was a greater cell integrity among germ cells that were not cryopreserved, based on criteria assessed (detachment of cells from basal membrane, retraction of seminiferous tubule epithelium, visibility of the spermatogonia nucleoli and nuclear spermatogonia condensation), for both sizes of fragments. The values for these variables decreased after cryopreservation, with there being no differences as a result of size of fragment stored or between cryoprotectants used (P > 0.05). The staining for caspase-3 differed for the cytoplasm, nuclei and germ cells. Assessment of these staining patterns indicated the fresh fragments had an amount of cell damage and there was a similar amount of damage detected in cryopreserved fragments. This finding indicated that there was considerable efficacy in preserving the tissue fragments with use of the freezing protocols that were evaluated in this study.

Clinical and pathological aspects of gossypiboma in a dog: case report / Aspectos clínico-patológicos do gossipiboma em cão: relato de caso

A female dog was treated at the Veterinary Hospital Governador Laudo Natel for symptomscaused by an intra-abdominal gossypiboma (gossypium: cotton; boma: place of hiding). Showing high morbidity and mortality, the gossypiboma is a granulomatous reaction that is formed in response to exposure to a textile matrix. The objective of this report is to describe the clinical and pathological findings of an intra-abdominal gossypiboma in a female dog. This patient arrived at urgent care showing increased abdominal volume, pain, and cachexia. Ultrasonography showed a regular contour formation that could not be completely delimited. The patient underwent exploratory laparotomy and died in the postoperative period. The gossypiboma had a smooth, firm, and purple-gray exterior surface. Microscopy revealed a severe reaction of desmoplasia around the necrotic region, residual textile material, and adipose tissue. Negligence by veterinarians leading to this condition can result in patients' death as well as causing psychological damage to owners.(AU)

Uma cadela foi atendida no Hospital Veterinário "Governador Laudo Natel", apresentando sinais clínicos oriundos de gossipiboma intra-abdominal (gossypium: algodão e boma: local de ocultação), tais como aumento de volume abdominal, regurgitação e caquexia. A ultrassonografia evidenciou formação de contorno regular que não podia ser totalmente delimitada. A paciente foi submetida à laparotomia exploratória e veio a óbito no período pós-operatório. O gossipiboma mostrou-se com superfície externa lisa, firme e roxo-acinzentada apresentando-se à microscopia como reação severa de desmoplasia ao redor de material necrótico, resíduos de material têxtil e do tecido adiposo. A negligência por parte dos médicos veterinários pode levar pacientes ao óbito, bem como trazer prejuízos psicológicos aos proprietários. Objetiva-se, com este relato, descrever os achados clínicos e patológicos de uma cadela que apresentou um gossipiboma intra-abdominal.(AU)

A importância dos exames neurológico e radiográfico no diagnóstico de lesões cervicais em equino: relato de caso / Fundamentals of neurological and radiographic examination on diagnosis of cervical lesions of a horse: case report

As lesões vertebrais cervicais são relativamente comuns nos equinos. No entanto, são poucas as descrições a respeito desse tema na literatura. Acredita-se que esse fato se deva, em parte, às limitações relacionadas ao diagnóstico. Em virtude de um traumatismo cervical, o animal pode apresentar sinais clínicos de paresia e paraplegia até tetraparesia ou tetraplegia, que podem culminar em estado comatoso e, eventualmente, em morte. Esses sinais clínicos podem ser detectados no exame físico, porém, quando associado à imaginologia (avaliação radiográfica), permite a obtenção de diagnóstico mais preciso da lesão. Consequentemente, um prognóstico mais adequado pode ser estabelecido para o paciente. Em face do exposto, apresenta-se o caso de uma égua Mangalarga, de quatro anos de idade, com sinais de tetraplegia pós-trauma, aumento de tônus muscular e paralisia espástica, com hiperreflexia, nos quatros membros. Por meio dos exames neurológicos realizados na paciente, em associação com as imagens radiográficas convencionais e contrastadas, foi possível estabelecer o diagnóstico do quadro. O animal apresentava lesões envolvendo as duas primeiras vértebras cervicais, atlas (C1) e áxis (C2), e a quinta e a sexta vértebra (C5-C6), destacando fratura de C1, deslocamento do dente de C2 e subluxação entre C5 e C6, com compressão ventrodorsal da medula espinhal. Confirmadas as suspeitas neurológicas e mediante prognóstico desfavorável, o animal foi submetido à eutanásia, com consentimento do proprietário. As lesões observadas nas imagens radiográficas foram confrontadas e confirmadas na necropsia.(AU)

Cervical vertebral lesions are relatively common in equine patients, but its diagnosis seems challenging for the clinician. Clinical signs most commonly associated with this clinical feature are paresis, paraplegia, tetraparesis, tetraplegia, which may lead the patient to a comatose state, and even death. Fortunately, these clinical signs can be perceived during physical examination. All of which, alongside imaging techniques, may allow a precise diagnose of the location of lesions and prediction of a prognosis for the patient. We present a case report of a Mangalarga mare, with four years of age, presenting signs of post-traumatic tetraplegia with spastic paralysis with increased muscular tonus and hyperreflexia in all four limbs. A thorough neurological examination of the patient associated with conventional and contrast medium radiographic imaging of the cervical region identified lesions at the atlas (C1) and axis (C2, and fifth (C5) and sixth (C6) cervical vertebrae. Fracture of the atlas vertebrae, dislocation of the axis' dens as well as subluxation of C5-C6 causing ventrodorsal medullary compression of the spinal cord were diagnosed radiographically. Due to the extent of the lesions the animal was euthanatized with the owner's consent. All lesions identified radiographically were posteriorly confirmed during post-mortem examination.(AU)

Epitelização de enxertos cutâneos em feridas recentes de coelhos tratados com membrana amniótica canina e/ou laserterapia / Skin graft epithelialization in rabbit fresh wounds treated with amniotic membrane and/or laser therapy

A enxertia cutânea é uma técnica cirúrgica simples e bastante útil para o reparo de feridas, principalmente aquelas onde existe dificuldade da aplicação do fechamento primário ou de outras técnicas reconstrutivas. Entretanto, para a sobrevivência do enxerto, é necessário que o leito da ferida esteja saudável e com presença de tecido de granulação exuberante. O objetivo do presente trabalho foi avaliar a aplicação da membrana amniótica e da laserterapia como potenciais estimulantes da cicatrização em enxertos aplicados em feridas sem tecido de granulação. Foram utilizados 42 coelhos, divididos em quatro grupos: grupo controle (GC), grupo membrana (GM), grupo laser (GL) e grupo membrana e laser (GML), submetidos a avaliações macro e microscópicas. Na avaliação macroscópica, foi possível notar que os pacientes dos grupos nos quais a membrana amniótica foi utilizada (GM e GML) apresentaram evidências associadas à acentuada reação inflamatória, à falha de integração do enxerto e à consequente necrose dele. Já os pacientes do GL apresentaram melhor aspecto do enxerto no último dia de avaliação. Na análise microscópica, observou-se intensa integração do enxerto à derme, reepitelização acentuada e escassas células inflamatórias no local do enxerto no GL. O contrário foi observado nos pacientes do GM e GML, nos quais aparentemente houve rejeição da membrana. A formação de colágeno não se correlacionou com outros fatores, como inflamação e necrose, em nenhum dos grupos de tratamento. Dessa forma, é possível afirmar que a laserterapia mostrou ser efetiva, contribuindo para o processo cicatricial e a integração do enxerto. Já a membrana amniótica canina não deve ser utilizada para esse fim, pois provoca intensa reação inflamatória, além de impedir a nutrição do enxerto.(AU)

Skin grafting is a simple surgical technique and useful to repair wounds, especially those where there is a difficulty to apply primary closure skin or other reconstructive techniques. However, for graft survival a healthy wound bed and the presence of an exuberant granulation tissue are necessary. This study aimed to evaluate the application of amniotic membrane and laser therapy as potential healing stimulants in grafts applied in wounds without granulation tissue. For this, we used 42 rabbits divided into four treatment groups, control group (CG), membrane group (MG), laser group (LG) and membrane and laser group (MLG), submitted to macroscopic and microscopic evaluation. The macroscopic examination showed that the patients of the groups where the amniotic membrane was used in order to stimulate re-epithelialization (MG and MLG) presented evidences associated with severe inflammatory reaction, graft integration failure and consequent necrosis. LG patients apparently had the best graph aspect in the last valuation date. Microscopic examination showed intense integration of the graft to the dermis, high re-epithelialization level, and scarce inflammatory cells in the graft site of LG patients. The opposite was observed in patients in the MG and MLG groups, where a rejection of the membrane was observed. Finally, collagen formation was not correlated with other factors such as inflammation and necrosis in any of the treatment groups. We can conclude that laser therapy was effective, contributing to the healing process and integration of the graft. Thus, canine amniotic membrane should not be used for this purpose because it causes intense inflammatory reaction besides avoiding graft nutrition.(AU)

Experimental infection of chickens by a flagellated motile strain of Salmonella enterica serovar Gallinarum biovar Gallinarum.

Salmonella enterica subsp. enterica serovar Gallinarum biovar Gallinarum (SG) causes fowl typhoid (FT), a septicaemic disease which can result in high mortality in poultry flocks. The absence of flagella in SG is thought to favour systemic invasion, since bacterial recognition via Toll-like receptor (TLR)-5 does not take place during the early stages of FT. In the present study, chicks susceptible to FT were inoculated with a wild type SG (SG) or its flagellated motile derivative (SG Fla(+)). In experiment 1, mortality and clinical signs were assessed, whereas in experiment 2, gross pathology, histopathology, systemic invasion and immune responses were evaluated. SG Fla(+) infection resulted in later development of clinical signs, lower mortality, lower bacterial numbers in the liver and spleen, and less severe pathological changes compared to SG. The CD8(+) T lymphocyte population was higher in the livers of chicks infected with SG at 4 days post-inoculation (dpi). Chicks infected with SG had increased expression of interleukin (IL)-6 mRNA in the caecal tonsil at 1 dpi and increased expression of IL-18 mRNA in the spleen at 4 dpi. In contrast, the CD4(+) T lymphocyte population was higher at 6 dpi in the livers of birds infected with SG Fla(+). Therefore, flagella appeared to modulate the chicken immune response towards a CD4(+) T profile, resulting in more efficient bacterial clearance from systemic sites and milder infection.

Cardiac lesions in 30 dogs naturally infected with Leishmania infantum chagasi.

The hearts of 30 dogs naturally infected with Leishmania infantum chagasi were evaluated histologically and immunohistochemically. Myocardial lesions were detected in all dogs, including lymphoplasmacytic myocarditis (27/30), myonecrosis (24/30), increased interstitial collagen (22/30), lepromatous-type granulomatous myocarditis (7/30), fibrinoid vascular change (3/30), and vasculitis (1/30). The parasite was detected in the hearts of 20 of 30 dogs. The number of parasitized cells correlated with the intensity of the inflammation and with the number of granulomas. The results indicate that cardiac lesions are prevalent in dogs with naturally occurring leishmaniasis even in the absence of clinical signs of cardiac disease.

Laparoscopic ovum collection in sheep: gross and microscopic evaluation of the ovary and influence on ooctye production.

Gross and microscopic evaluations of the ovaries of sheep subjected to successive ovum collection were performed with the objective of identifying if there was interference in oocyte production and morphology of gonads. Gross evaluation of the internal genital tract was also performed. Eighteen ewes of the Santa Inês breed were randomly distributed into three experimental groups of six animals each; G0, G1 and G9 with no, one and nine repetitions, respectively. Estrous synchronization was achieved with a short protocol using MAP followed by single dose stimulation with 80mg of FSHp and 300 IU of eCG (IM). Laparoscopic ovum pick-up was performed 36h later, with 7-day intervals. The number of visualized follicles, aspirated follicles and oocytes recovery was recorded. After the last intervention, ovariectomy was performed for evaluation of gross and microscopical appearance and existence of lesions caused by follicular puncture, which were classified as absent (0), mild (1), moderate (2) and severe (3). The number of visualized follicles, aspirated follicles and oocytes recovery were 13.2±2.0, 11.3±3.0 and 5.8±2.3, respectively, with a recovery rate of 51.7%. No statistical difference was found between the nine sessions (P>0.05). Nine sessions of superovulation and ovum collection procedures did not cause ovarian lesions and did not interfere with the production of follicles in ewes of the Santa Inês breed.

Morphological abnormalities and apoptosis in lamellar tissue of equines after intestinal obstruction and treatment with hydrocortisone / Alterações morfológicas e apoptose em tecido laminar de equinos após obstrução intestinal e tratamento com hidrocortisona

Four experimental groups of equines were used in order to study morphological abnormalities and apoptosis in lamellar tissue. Group Cg (control) was composed of animals without any surgical procedure; group Ig (instrumented), animals that underwent enterotomy; group Tg (treated), animals that were subjected to intestinal obstruction and were treated with hydrocortisone; and group Ug (untreated), animals that were subjected to intestinal obstruction without treatment. The lamellar tissue was analyzed regarding the presence of tissue abnormalities and apoptosis. No morphological abnormalities were observed in animals of surgical groups, and no difference in apoptosis was observed between groups. It was concluded that intestinal obstruction allowed laminitis to develop, probably by systemic activation, and that the maneuvers performed in the enterotomy aggravated the process. Hydrocortisone did not aggravate the lesions of the lamellar tissue.

Foram utilizados quatro grupos de equinos para estudar alterações morfológicas e apoptose no tecido lamelar. O grupo CG (controle) foi composto por animais sem o procedimento cirúrgico; o grupo Ig (instrumentado), por animais submetidos à enterotomia; o grupo Tg (tratado), por animais submetidos à obstrução intestinal e tratados com hidrocortisona; e o grupo Ug (não tratado), por animais submetidos à obstrução intestinal, sem tratamento. O tecido laminar foi analisado quanto à presença de alterações morfológicas e de apoptose. Foram observadas alterações morfológicas nos equinos submetidos a procedimento cirúrgico, mas não houve diferença entre os grupos com relação às células apoptóticas. Conclui-se que a obstrução intestinal permite o desenvolvimento da laminite provavelmente por ativação sistêmica e que as manobras realizadas na enterotomia podem ser consideradas como agravantes no processo. A hidrocortisona não agravou as lesões do tecido laminar.

Leukocyte entry into the CNS of Leishmania chagasi naturally infected dogs.

In dogs, there is an association of chronic visceral leishmaniasis with neurological symptoms, and very few publications have investigated whether these neurological manifestations correlate with specific alterations in brain. A total of 42 mixed-breed adult dogs were selected from the Veterinary Hospital of UNESP-Araçatuba and the Control Zoonosis Center in Araçatuba, São Paulo State, Brazil, which is an endemic area for visceral leishmaniasis. Animals presenting positive ELISA and/or positive parasitological diagnosis of Leishmania were enrolled in the group of infected dogs (n=32). Animals with negative ELISA results and parasitological tests for Leishmania, including a negative immunofluorescence test for toxoplasmosis and neosporosis, were included as the control group (n=10). Brain samples were collected, stored in 10% buffered formalin and subjected to routine histological procedures, following by staining with haematoxylin-eosin (HE) and immunohistochemical examination for T and B lymphocytes and phagocytic cells. Cerebrospinal fluid was collected to determine the anti-Leishmania antibody titers. Histological examination of HE stains demonstrated intense inflammatory infiltrate, primarily in the choroid plexus, which was composed of mononuclear cells with no detectable parasites. Immunohistochemistry revealed that CD3(+) T lymphocytes were the major components of the inflammatory infiltrate at the choroid plexus and in the brain. Infected dogs had more CD3(+) T cells than uninfected animals (P=0.0002). Cerebrospinal fluid from infected dogs contained high titers of anti-Leishmania antibodies in comparison with control animals (P<0.0001), which suggests a compromise of the blood-cerebrospinal fluid barrier. Leukocyte entry into the brain suggests the participation of these cells in the pathogenesis of neurological disorders during the advanced stages of leishmaniasis and confirms that the choroid plexus is an important structure for T cell influx.