Expression of matrix metalloproteinase-2 and metalloproteinase-9 in the skin of dogs with visceral leishmaniasis.

The skin is the first organ to be infected by the parasite in canine visceral leishmaniasis. The enzyme matrix metalloproteinase (MMP) acts towards degradation of the extracellular matrix (ECM) and modulation of the inflammatory response against many kinds of injuries. The aims of this study were to evaluate the expression of MMP-2 and MMP-9 through immunohistochemistry and zymography on the skin (muzzle, ears, and abdomen) of dogs that were naturally infected by Leishmania spp. and to compare these results with immunodetection of the parasite and with alterations to the dermal ECM. Picrosirius red staining was used to differentiate collagen types I and III in three regions of the skin. The parasite load, intensity of inflammation, and production of MMP-2 (latent) and MMP-9 (active and latent) were higher in the ear and muzzle regions. MMP-9 (active) predominated in the infected group of dogs and its production was significantly different to that of the control group. Macrophages, lymphocytes, and plasma cells predominated in the dermal inflammation and formed granulomas in association with degradation of mature collagen (type I) and with discrete deposition of young collagen (type III). This dermal change was more pronounced in dogs with high parasite load in the skin. Therefore, it was concluded that the greater parasite load and intensity of inflammation in the skin led consequently to increased degradation of mature collagen, caused by increased production of MMPs, particularly active MMP-9, in dogs with visceral leishmaniasis. This host response profile possibly favors systemic dissemination of the parasite.

Immunodetection of cells with a CD44+/CD24- phenotype in canine mammary neoplasms.

BACKGROUND: Cancer stem cells (CSCs) are able to self-renew and to form metastases. Using flow cytometry, CSCs were detected in canine mammary tumors as cells CD44(+) and CD24(-). The aim of this study was to detect these CSCs by immunohistochemistry and correlate their frequency with canine mammary neoplasm grade and histopathological type.130 mammary neoplasm samples were selected from tissue blocks at the Department of Pathology at UNESP and classified according to (BJVP 4:153-180, 2011). These samples were composed by adenomas, lymph node metastases, solid carcinomas grades II and III, tubular, papillary and carcinomas in mixed tumor grades I, II and III. Immunohistochemistry was performed with antibodies against CD44 and CD24. Linear regression was performed using Pearson's correlation test. RESULTS: The value at CD44 was positive and CD24 becomes zero was 46.75%. Cells with a CD44(+)/CD24(-) phenotype were detected in 40 out of 130 samples with an advantage of high grade tumors (II and III) and metastases among tubular, papillary and carcinomas in mixed tumors. In these samples, percentages of cells stained by CD44 and CD24 antibodies were 62.2% and 0%, respectively. Published reports usually correlate grade III tumors with the expression of CD44 but not with CD24 expression. Studies using flow cytometry have found CSC frequencies similar to those found in our study. CONCLUSIONS: Immunohistochemistry was found to be a reliable technique for the detection of CSCs in canine mammary neoplasms, and the frequency of these cells positively correlates with grades II and III tumors (poor prognosis).

Influence of serum progesterone levels on the inflammatory response of female dogs with visceral leishmaniosis.

The aim of this study was to evaluate the histopathological changes to the mammary gland that occur in female dogs with visceral leishmaniosis and to correlate the findings with the parasite load, inflammatory cell profile in mammary tissue and serum progesterone levels. For this, 20 adult female dogs that were naturally infected with Leishmania infantum, not spayed, not pregnant and free from mammary tumors were used. They were divided into two groups: G1 (n = 9) with high serum progesterone levels and G2 (n = 11) with low serum progesterone levels. The parasite load and the immunophenotype of leukocytes infiltrated into the mammary tissue (CD3, CD4, CD8 and MCA874) were evaluated using the immunohistochemical technique. In the mammary gland, chronic inflammatory infiltrate was mainly found in G1, sometimes associated with granulomatous inflammation, higher parasite load and higher density of cells immunolabeled for CD3, CD4, CD8 and MCA874. A significant positive correlation (p < 0.05) was observed between the parasite load and the immunolabeled leukocytes. The influence of the serum progesterone level in the mammary gland of infected female dogs can contribute to the maintenance of an immunosuppressive cell profile and favor the persistence of the parasite in this site.

Immune response pattern of the popliteal lymph nodes of dogs with visceral leishmaniasis.

The present study aimed to estimate the cell response and parasite load in the popliteal lymph nodes of dogs with visceral leishmaniasis (VL), comparing these findings with the clinical staging of the disease. From the necropsy, 33 dogs were classified as symptomatic (S), asymptomatic (A), or oligosymptomatic (O). Cytology and histopathology were used to determine any presence of microscopic lesions and immunohistochemistry, for parasite load. Dog hyperimmune serum was used as the primary antibody. The inflammatory infiltrate in lymph nodes consisted of macrophages and plasmocytes. The granulomas invaded the trabecular and sinusoid regions and sometimes compressed the lymphocytes of the cortical region (atrophy) and medullary cord cells. Parasite load intensity was unrelated to the density of the macrophages infiltrating the lymph node. Significant differences in parasite load (P < 0.05) were observed between the three groups of infected dogs. Follicular hyperplasia of the cortical region occurred among A and O, while follicular atrophy predominated among S. The parasite load was the greatest among S, followed by O. It can be concluded that, regardless of clinical condition, the most evident cell response consisted of macrophages and plasmocytes. Lymphoid atrophy was observed among animals with intense granulomatous reaction and high parasite load, such as among the symptomatic dogs (P < 0.05). Likewise, the oligosymptomatic dogs also presented high density of parasites in the lymph nodes. Thus, we can confirm that dogs with clinical manifestations of VL have an immune system that is less effective for controlling infection by Leishmania chagasi, thereby favoring parasite multiplication.

Polarized M2 macrophages in dogs with visceral leishmaniasis.

The objective of the present study was to analyze the skin (nasal surface and ear regions), lymph nodes (popliteal and pre-scapular), spleen and liver of dogs with visceral leishmaniasis (VL), in order to investigate the relationship between the parasite load measured as DNA copy number of Alpha gene of DNA polymerase of Leishmania infantum by quantitative PCR and the number of M2 macrophages by immunohistochemistry. A set of 29 naturally infected dogs from an endemic area for VL were sampled and another set of six dogs negative for VL and from a non-endemic area were analyzed as the control group (C). The spleen presented the highest number of Leishmania DNA copies, with significant differences between the groups G1 and G2 (with and without skin lesions, respectively). The M2 phenotype immunostaining predominated among the macrophages in granulomas and inflammatory infiltrates of samples from the skin, lymph nodes and spleens examined. The presence of M2 macrophages in dogs from infected group differed significantly from the control group, in all organs analyzed, excepted liver. The highest proportion of M2 macrophages coincided with the highest parasitism loads found in more susceptible organs of VL dogs, even in the skin, considered a more resistant organ, while the liver showed low parasitism load and low immunostaining for M2 macrophages with no significant differences between infected and negative groups. It was concluded that the predominance of M2 phenotype in VL dogs favored the multiplication of Leishmania infantum in organs of dogs that are more susceptible to Leishmania infection, as skin, lymph nodes and spleen.

Influence of apoptosis on liver and spleen resistance in dogs with visceral leishmaniosis.

The aim of this study was to evaluate apoptosis and parasite load in the liver and spleen of dogs with visceral leishmaniosis (VL), using immunohistochemistry. Liver and spleen samples from 71 dogs with VL were used. The parasite load in the spleen and liver showed significant difference between organs in infected group (P=0.0219). The density of the parasite load in the spleen (median=2.4) was higher than liver (median=0.8). Immunodetection of apoptotic cells was predominant in lymphocytes and differ between the infected and control group in spleen (P=0.0307) and liver (P=0.0346). There was a significant correlation between apoptosis and parasite load (P = 0.0084; r=0.3104) only in the spleen of the infected group, where it was observed that, when increasing the number of apoptotic cells increases the parasitic load. It was concluded that the liver and spleen of infected dogs presented greater numbers of cells undergoing apoptosis (lymphocytes) than the control group, thus suggesting that this process may be contributing towards the survival of Leishmania in these organs, because lymphocyte in apoptosis did not have the ability to present and recognize the antigen, allowing the survival of the parasite.

Relationship between the inflammatory infiltrate and the degree of differentiation of the canine cutaneous squamous cell carcinoma.

The inflammatory response may have pro or anti-neoplastic effects in tumors, depending on the histological type and malignancy level of the tumor. This study aimed to evaluate the profile of predominant inflammatory cells by immunohistochemistry in cutaneus squamous cell carcinoma (SCC) of dogs, comparing it with the degree of differentiation of the tumor. Twenty samples of SCC were analyzed. The tumors were histologically classified into two groups, differentiated SCC (SCCd=12) and undifferentiated SCC (SCCu=08). The tumor inflammatory infiltrate was determined by immunohistochemistry, in order to identify macrophages, lymphocytes and plasma cells. The comparison between groups, SCCd and SCCu, was not significant concerning the density of macrophages (P=1.0), T lymphocytes (P=0.335) and plasma cells (P=0.075). However, when comparing the inflammatory infiltrate in each group, the macrophages were the predominant cell type in both groups, a significant difference was found in the SCCd with plasma cells (P<0.0001). In the SCCu the difference occurred with lymphocytes (P<0.05) and plasma cells (P=0.0006). It could be concluded that the presence of inflammation in cases of SCC does not play a role in the differentiation of the neoplasm, since the inflammatory infiltrate was similar in both groups of SCC.

Morphological changes in the bone marrow of the dogs with visceral leishmaniasis.

The aim of this study was to evaluate the most frequent lesions in the bone marrow of dogs naturally infected by Leishmania (Leishmania) chagasi. Thirty-three dogs sacrificed at the Zoonosis Control Center of Araçatuba, a municipality endemic for visceral leishmaniasis (VL), were used. The animals were classified as asymptomatic, oligosymptomatic, and symptomatic groups. At the necropsy, bone marrow samples were collected from the femur, fixed, processed, and stained with hematoxylin and eosin. The lesion intensity was classified as mild, moderate, or severe. The parasite load was determined using immunohistochemistry. The most important lesions consisted of multifocal to diffuse granulomas, megakaryocytic dysplasia, and medullary aplasia. There were no statistical differences between the three clinical groups regarding parasite load and lesion intensity. Asymptomatic dogs also presented high parasitism in the bone marrow as dogs with clinical signs of VL. It was concluded that, regardless of clinical group, the bone marrow is a site for multiplication of Leishmania chagasi. Possibly, the bone marrow dysplasia may arise from the presence of many parasitized and activated macrophages in this organ. Consequently, it affects the profile of hematopoietic cells in the bone marrow and systemic circulation.

Morphological changes and parasite load of the adrenal from dogs with visceral leishmaniasis.

The objective of this study was to analyze morphological changes and parasite loads in the adrenal gland from 45 dogs with visceral leishmaniasis (VL). The animals were from the Zoonosis Control Center of Araçatuba, state of São Paulo, which is an endemic region for the disease. These animals were euthanized due to positive diagnoses of VL. The dogs were classified into asymptomatic, oligosymptomatic and symptomatic groups. The parasite load was determined by immunohistochemistry, using VL-positive dog hyperimmune serum. Nine dogs showed an inflammatory infiltrate composed, predominantly, of plasma cells and macrophages. However, only eight dogs showed macrophages with amastigote forms of the parasite, immunolabeled in the cytoplasm. The medullary and reticular layers were the most affected areas, possibly due to a favorable microenvironment created by hormones in these regions. The density of parasites in the glandular tissue was not associated with clinical signs of VL (P > 0.05). However, the presence of the parasite was always associated with the presence of a granulomatous inflammatory infiltrate. This gland may not be an ideal place for the parasite's multiplication, but the presence of injuries to the glandular tissue could influence the dog's immune system, thus favoring the parasite's survival in the host's different organs.

Influence of apoptosis on the cutaneous and peripheral lymph node inflammatory response in dogs with visceral leishmaniasis.

In canine visceral leishmaniasis (CVL), the abnormalities most commonly observed in clinical examination on the animals are lymphadenomegaly and skin lesions. Dogs are the main domestic reservoir for the protozoon Leishmania (L.) chagasi and the skin is the main site of contamination by the vector insect. Some protozoa use apoptosis as an immunological escape mechanism. The aim of this study was to correlate the presence of apoptosis with the parasite load and with the inflammatory response in the skin and lymph nodes of dogs naturally infected with Leishmania (L.) chagasi. Thirty-three dogs from the municipality of Araçatuba (São Paulo, Brazil) were used, an endemic area for CVL. Muzzle, ear and abdominal skin and the popliteal, subscapular, iliac and mesenteric lymph nodes of symptomatic (S), oligosymptomatic (O) and asymptomatic (A) dogs were analyzed histologically. The parasite load and percentage apoptosis were evaluated using an immunohistochemical technique. Microscopically, the lymph nodes presented chronic lymphadenitis and the skin presented plasmacytic infiltrate and granulomatous foci in the superficial dermis, especially in the ear and muzzle regions. The inflammation was most severe in group S. The parasite load and apoptotic cell density were also greatest in this group. The cause of the lymphoid atrophy in these dogs was correlated with T lymphocyte apoptosis, thus leaving the dogs more susceptible to CVL. The peripheral lymph nodes presented the greatest inflammatory response. Independent of the clinical picture, the predominant inflammatory response was granulomatous and plasmacytic, both in the skin and in the peripheral lymph nodes. The ear skin presented the greatest intensity of inflammation and parasite load, followed by the muzzle skin, in group S. The ear skin area presented a non-significant difference in cell profile, with predominance of macrophages, and a significant difference from group A to groups O and S. It was seen that in these areas, there were high densities of parasites and cells undergoing apoptosis, in group S. The association between apoptosis and parasite load was not significant in the lymph nodes, but in the muzzle regions and at the ear tips, a positive correlation was seen between the parasite load and the density of cells undergoing apoptosis. The dogs in group S had the highest parasite load and the greatest number of apoptotic cells, thus suggesting that the parasite had an immune evasion mechanism, which could be proven statistically in the skin.

Influence of apoptosis on liver and spleen resistance in dogs with visceral leishmaniosis / Influência da apoptose na resistência hepática e esplênica de cães com leishmaniose visceral

Abstract The aim of this study was to evaluate apoptosis and parasite load in the liver and spleen of dogs with visceral leishmaniosis (VL), using immunohistochemistry. Liver and spleen samples from 71 dogs with VL were used. The parasite load in the spleen and liver showed significant difference between organs in infected group (P=0.0219). The density of the parasite load in the spleen (median=2.4) was higher than liver (median=0.8). Immunodetection of apoptotic cells was predominant in lymphocytes and differ between the infected and control group in spleen (P=0.0307) and liver (P=0.0346). There was a significant correlation between apoptosis and parasite load (P = 0.0084; r=0.3104) only in the spleen of the infected group, where it was observed that, when increasing the number of apoptotic cells increases the parasitic load. It was concluded that the liver and spleen of infected dogs presented greater numbers of cells undergoing apoptosis (lymphocytes) than the control group, thus suggesting that this process may be contributing towards the survival of Leishmania in these organs, because lymphocyte in apoptosis did not have the ability to present and recognize the antigen, allowing the survival of the parasite.

Resumo O objetivo deste estudo foi avaliar a apoptose e a carga parasitária no fígado e baço de cães com leishmaniose visceral (LV), pela técnica de imuno-histoquímica. Foram utilizadas amostras de fígado e baço de 71 cães com LV. A carga parasitária no baço e fígado mostrou diferença significativa entre os órgãos no grupo infectado (P=0,0219). A densidade da carga de parasita no baço (média=2,4) foi maior do que no fígado (média=0,8). A imunodetecção de células em apoptose foi predominante nos linfócitos, com diferenças entre o grupo infectado e controle no baço (P=0,0307) e fígado (P=0,0346). Houve uma correlação positiva fraca entre apoptose e carga parasitária (P=0,0084; r=0,3104) apenas no baço do grupo infectado, onde observou-se que quando aumentava o número de células em apoptose aumentava a carga parasitária. Concluiu-se que o fígado e o baço de cães infectados apresentam um maior número de células que sofrem apoptose (linfócitos) do que o grupo controle, sugerindo que este processo possa contribuir para a sobrevivência de Leishmania nestes órgãos, pois os linfócitos em apoptose não tiveram a capacidade de apresentar e reconhecer o antígeno, permitindo a sobrevivência do parasita.

Influence of apoptosis on liver and spleen resistance in dogs with visceral leishmaniosis / Influência da apoptose na resistência hepática e esplênica de cães com leishmaniose visceral

Abstract The aim of this study was to evaluate apoptosis and parasite load in the liver and spleen of dogs with visceral leishmaniosis (VL), using immunohistochemistry. Liver and spleen samples from 71 dogs with VL were used. The parasite load in the spleen and liver showed significant difference between organs in infected group (P=0.0219). The density of the parasite load in the spleen (median=2.4) was higher than liver (median=0.8). Immunodetection of apoptotic cells was predominant in lymphocytes and differ between the infected and control group in spleen (P=0.0307) and liver (P=0.0346). There was a significant correlation between apoptosis and parasite load (P = 0.0084; r=0.3104) only in the spleen of the infected group, where it was observed that, when increasing the number of apoptotic cells increases the parasitic load. It was concluded that the liver and spleen of infected dogs presented greater numbers of cells undergoing apoptosis (lymphocytes) than the control group, thus suggesting that this process may be contributing towards the survival of Leishmania in these organs, because lymphocyte in apoptosis did not have the ability to present and recognize the antigen, allowing the survival of the parasite.

Resumo O objetivo deste estudo foi avaliar a apoptose e a carga parasitária no fígado e baço de cães com leishmaniose visceral (LV), pela técnica de imuno-histoquímica. Foram utilizadas amostras de fígado e baço de 71 cães com LV. A carga parasitária no baço e fígado mostrou diferença significativa entre os órgãos no grupo infectado (P=0,0219). A densidade da carga de parasita no baço (média=2,4) foi maior do que no fígado (média=0,8). A imunodetecção de células em apoptose foi predominante nos linfócitos, com diferenças entre o grupo infectado e controle no baço (P=0,0307) e fígado (P=0,0346). Houve uma correlação positiva fraca entre apoptose e carga parasitária (P=0,0084; r=0,3104) apenas no baço do grupo infectado, onde observou-se que quando aumentava o número de células em apoptose aumentava a carga parasitária. Concluiu-se que o fígado e o baço de cães infectados apresentam um maior número de células que sofrem apoptose (linfócitos) do que o grupo controle, sugerindo que este processo possa contribuir para a sobrevivência de Leishmania nestes órgãos, pois os linfócitos em apoptose não tiveram a capacidade de apresentar e reconhecer o antígeno, permitindo a sobrevivência do parasita.

Morphological changes and parasite load of the adrenal from dogs with visceral leishmaniasis / Alterações morfológicas e carga parasitária da adrenal de cães com leishmaniose visceral

The objective of this study was to analyze morphological changes and parasite loads in the adrenal gland from 45 dogs with visceral leishmaniasis (VL). The animals were from the Zoonosis Control Center of Araçatuba, state of São Paulo, which is an endemic region for the disease. These animals were euthanized due to positive diagnoses of VL. The dogs were classified into asymptomatic, oligosymptomatic and symptomatic groups. The parasite load was determined by immunohistochemistry, using VL-positive dog hyperimmune serum. Nine dogs showed an inflammatory infiltrate composed, predominantly, of plasma cells and macrophages. However, only eight dogs showed macrophages with amastigote forms of the parasite, immunolabeled in the cytoplasm. The medullary and reticular layers were the most affected areas, possibly due to a favorable microenvironment created by hormones in these regions. The density of parasites in the glandular tissue was not associated with clinical signs of VL (P > 0.05). However, the presence of the parasite was always associated with the presence of a granulomatous inflammatory infiltrate. This gland may not be an ideal place for the parasite's multiplication, but the presence of injuries to the glandular tissue could influence the dog's immune system, thus favoring the parasite's survival in the host's different organs.

O objetivo deste estudo foi analisar as alterações morfológicas e a carga parasitária da glândula adrenal de 45 cães com leishmaniose visceral (LV). Os animais eram provenientes do Centro de Controle de Zoonoses (CCZ) de Araçatuba (SP), região endêmica para a doença. Esses animais são submetidos à eutanásia, devido ao diagnóstico positivo para LV. Os cães foram classificados nos grupos assintomático, oligossintomático e sintomático. A determinação da carga parasitária foi feita por imuno-histoquímica, com utilização de soro hiperimune de cão positivo para LV. Em nove cães, verificou-se um infiltrado inflamatório, composto predominantemente por plasmócitos e macrófagos. Entretanto, apenas oito cães apresentaram macrófagos com formas amastigotas do parasito imunomarcadas em seu citoplasma. As camadas medular e reticulada foram as mais afetadas, possivelmente por um microambiente favorável criado pelos hormônios nestas regiões. A densidade de parasitos no tecido glandular não foi relacionada com os sinais clínicos de LV (P > 0,05). No entanto, a presença do parasito sempre esteve associada à presença de infiltrado inflamatório granulomatoso. Possivelmente, essa glândula não é um sítio ideal para a multiplicação do protozoário, mas a presença de injúrias no tecido glandular poderia influenciar o sistema imune do cão, favorecendo a sobrevivência do parasito nos diferentes órgãos do hospedeiro.

Arranjos configurados pelos nervos do plexo braquial no urubu (Coragyps atratus foetens -Linnaeus, 1758) / The arragement of configurat of nervs of brachial plexus in vulture (Coragyps atratus foetens - Linnaeus, 1758)

O estudo anatômico da origem e distribuição do plexo braquial no urubu (Coragyps atratus foetens) foi efetuado em 14 animais, adultos,machos e fêmeas oriundos da região de Araçatuba (SP). Após a fixação em solução aquosa de formaldeído a 10,00%, realizou-se a dissecação bilateral da origem dos plexos braquiais e sua distribuição. A análise permitiu verificar, que o plexo braquial do urubu, possui quatro troncos, os quais originam-se dos ramos ventrais dos nervos espinhais cervicais onze (C11), doze (C12) e treze (C13) e dos ramos ventrais dos nervos espinhais torácicos um (T1) e (T2) em 100% dos casos em ambos antímeros. O primeiro tronco parte de C11 nos antímeros direito e esquerdo, e da margem cranial do cordão dorsal partem os nervos subescapular, subcoracoescapular e supracoracóide; em ambos antímeros (100%). O segundo tronco origina-se de C12 em ambos antímeros, dando origem ao cordão dorsal, dele partem os nervos axilar, radial e anconeal (100%). O terceiro tronco do plexo origina-se unicamente de C13 (100%) e o quarto tronco de T1 em ambos antímeros (100%). Os troncos de C12, C13 e T1 unem-se dando origem ao cordão ventral, o qual partem os nervos medianoulnar quese divide em mediano e ulnar e os nervos peitorais (100%). T2 emite filamentos que se unem à raiz nervosa de T1, sendo um filamento(35,55%) e dois filamentos (64,29%) no antímero direito, enquanto que no esquerdo um filamento (42,85%), dois filamentos (50%) e três filamentos (7,15%)

The anatomical study of the origin and distribution of brachial plexus in vulture (Coragyps atratus foetens) was performed in 14 animals, adults,males and females, deriving of the region of Araçatuba (SP). After fixed in watery solution of formaldehyde 10,00%, became dissected bilateral of the origin of the brachial plexus, and yours distribution. The analysis allowed verify that brachial plexus of vulture, four root possess, which originate from the ventral branches of cervical spinal nerves eleven (C11),twelve (C12) and thirteen (C13) and of the ventral branches of thoracic spinal nerve one (T1) and (T2) in both sides (100%). The first root part of C11 in the right and left side, and of the edge skull of the dorsal lace the nerves subescapular, subcoracoescapular e supracoracóide in bothantímeros (100%). As the root originates from C12 in both sides, giving origin to the dorsal lace, giving the axilar, radial and anconeal nerves(100%). The third root of plexus originates solely from C13 (100%)and the root of T1 in both sides (100%). The roots of C12, C13 and T1 was united the ventral lace originates, which breaks the nerves to medianoulnar that divided in medium and to ulnar and the pectoral nerves (100%). T2 emits filaments that join it root nervous of T1,being one filament (35,55%) and two filaments (64,29%) in side right,while that in the left one filament (42,85%), two filaments (50%) and three filaments (7,15%)

Ramificação e distribuição dos nervos frênicos no músculo diafragma do gato doméstico / Ramification and distribution of the phrenic nerves in the muscle diaphragm of the domestic cat

A ramificação e a distribuição dos nervos frênicos direito e esquerdo foram estudadas em 30 músculos diafragmas de gatos domésticos sem raça definida, 7 machos e 23 fêmeas, conservados em solução de formaldeído a 10 %. Mediante fixação e dissecção, observou-se que os nervos frênicos se ramificaram para as respectivas partes carnosas do músculo diafragma, pars lumbalis, costalis e sternalis, e terminaram em maior freqüência em tronco dorsolateral e ramo ventral (63,33 %). Notou-se ainda os seguintes arranjos: ramos dorsal, lateral e ventral (25,0 %); ramo dorsal e tronco ventrolateral (6,66 %); troncos dorsolateral e ventrolateral (3,33 %); tronco dorsolateral, ramos lateral e ventral (1,66 %). Os nervos frênicos se distribuíram simetricamente em 11 exemplares (36,66 %), exibindo somente a terminação em tronco dorsolateral e ramo ventral. Os ramos dorsais inervaram a pars lumbalis (73,33 % à direita e 56,66 % à esquerda) e a pars costalis (13,33 % à direita e 10,0 % à esquerda). O ramo dorsal direito supriu a crus mediale dexter do pilar direito (100,0 %) e o ramo dorsal esquerdo inervou a crus mediale sinister do pilar direito e o pilar esquerdo (100,0 %). Os ramos laterais inervaram a pars lumbalis (23,33 % à direita e 33,33 % à esquerda), a pars costalis (96,66 % à direita e 100,0 % à esquerda) e a pars sternalis (3,33 % somente à direita). Os ramos ventrais inervaram a região ventral da pars costalis (46,66 % à direita e 43,33 % à esquerda) e a pars sternalis (96,66 % à direita e 100,0 % à esquerda). Quatro animais fêmeas (13,33 %) exibiram cruzamento de fibras provenientes do ramo ventral esquerdo para o antímero direito sendo que em um desses exemplares (3,33 %) ocorreu conexão entre o ramo ventral esquerdo e o direito.

The ramification and the distribution of the phrenic nerves right and left had been studied in 30 muscles diaphragms of unknown breed adult domestic cats, 7 males and 23 females, and were fixed in 10 % buffered formalin solution. After fixation and dissection, it was observed that the phrenic nerves ramified for the respective carnous parts of the muscle diaphragm, pars lumbalis, costalis and sternalis, and had finished in higher frequency in dorsolateral trunk and ventral branch (63.33 %). We observed the following arrangements: dorsal, ventral and lateral branches (25.0 %); dorsal branch and ventrolateral trunk (6.66 %); dorsolateral and ventrolateral trunks (3.33 %); dorsolateral trunk, lateral and ventral branches (1.66 %). The phrenic nerves had distributed symmetrically in 11 samples (36.66 %), only showing the termination in dorsolateral trunk and ventral branch. The dorsal branches supplied pars lumbalis (73.33 % to right and 56.66 % to the left) and pars costalis (13.33 % to right and 10.0 % to the left). The right dorsal branch supplied the crus mediale dexter of the right pillar (100.0 %) and the left dorsal branch supplied the crus mediale sinister of the right pillar and the left pillar (100.0 %). The lateral branches supply pars lumbalis (23.33 % to right and 33.33 % to the left), pars costalis (96.66 % to right and 100.0 % to the left) and pars sternalis (3.33 % only to the right). The ventral branches supplied the ventral region of pars costalis (46.66 % to right and 43.33 % to the left) and pars sternalis (96.66 % to right and 100.0 % to the left). Four female animals (13.33 %) had shown fibers crossing proceeding from the left ventral branch for right antimere had been that in one of these samples (3.33 %) occurred connection between the left ventral branch and the right.