Tuberculosis caused by Mycobacterium caprae in a camel (Camelus dromedarius).

BACKGROUND: Animal tuberculosis (TB) is distributed worldwide and has a wide range of wild and domestic reservoirs. Few studies concerning TB in camelids have been published in the last decade, particularly as regards Old World Camelids (OWC), but the increase in reports of TB outbreaks in these species in recent years suggests a high susceptibility to the infection. CASE PRESENTATION: We studied a dromedary camel (Camelus dromedarius) herd (n = 24) in which a Mycobacterium caprae infection was detected. The TB infection was confirmed in one animal at necropsy through the detection of TB lesions, mainly in the abdominal organs, and the subsequent isolation of M. caprae (SB0157 spoligotype). The whole herd was additionally tested using cellular and humoral based diagnostic techniques. The intradermal tuberculin test results were compared with those obtained using P22 ELISA for the detection of specific antibodies against the M. tuberculosis complex. The TB infected animal was a positive reactor to both the intradermal tuberculin tests and P22 ELISA, while the others were negative to all the diagnostic tests. CONCLUSION: The present study found M. caprae infection in OWC. This is the first report of M. caprae infection in an OWC not living in a zoo. Since the animal was born in the herd and fed with goat's milk, this practice was suspected to be the potential source of TB infection, which was not confirmed in the other animals present in the herd. Moreover, our results highlight that the intradermal tuberculin test and the P22 ELISA could be valuable tools for the diagnosis of TB in OWC.

Evaluation of the Mycobacterium tuberculosis SO2 vaccine using a natural tuberculosis infection model in goats.

The development of new vaccines against animal tuberculosis (TB) is a priority for improving the control and eradication of this disease, particularly in those species not subjected to compulsory eradication programmes. In this study, the protection conferred by the Mycobacterium tuberculosis SO2 experimental vaccine was evaluated using a natural infection model in goats. Twenty-six goats were distributed in three groups: (1) 10 goats served as a control group; (2) six goats were subcutaneously vaccinated with BCG; and (3) 10 goats were subcutaneously vaccinated with SO2. Four months after vaccination, all groups were merged with goats infected with Mycobacterium bovis or Mycobacterium caprae, and tested over a 40 week period using a tuberculin intradermal test and an interferon-γ assay for mycobacterial reactivity. The severity of lesions was determined at post-mortem examination and the bacterial load in tissues were evaluated by culture. The two vaccinated groups had significantly lower lesion and bacterial culture scores than the control group (P<0.05); at the end of the study, the SO2 vaccinated goats had the lowest lesion and culture scores. These results suggest that the SO2 vaccine provides some protection against TB infection acquired from natural exposure.

Dietary vitamin A restriction affects adipocyte differentiation and fatty acid composition of intramuscular fat in Iberian pigs.

The aim of this study was to investigate whether dietary vitamin A level is associated with differences in adipocyte differentiation or lipid accumulation in Iberian pigs at early growing (35.8kg live weight) and at finishing (158kg live weight). Iberian pigs of 16.3kg live weight were allocated to two feeding groups, one group received 10,000IU of vitamin A/kg diet (control); the other group received a diet with 0IU of vitamin A (var) for the whole experimental period. The dietary vitamin A level had no effect on growth performance and carcass traits. The early suppression of vitamin A increased the preadipocyte number in Longissimus thoracis (LT) muscle in the early growth period (P<0.001) and the neutral lipid content and composition (higher MUFA and lower SFA content) at the end of the finishing period (P<0.05). Vitamin A restriction in young pigs increases their lipogenic potential without affecting carcass traits.

Distinguishing Intestinal Lymphoma From Inflammatory Bowel Disease in Canine Duodenal Endoscopic Biopsy Samples.

Inflammatory bowel disease (IBD) and intestinal lymphoma are intestinal disorders in dogs, both causing similar chronic digestive signs, although with a different prognosis and different treatment requirements. Differentiation between these 2 conditions is based on histopathologic evaluation of intestinal biopsies. However, an accurate diagnosis is often difficult based on histology alone, especially when only endoscopic biopsies are available to differentiate IBD from enteropathy-associated T-cell lymphoma (EATL) type 2, a small cell lymphoma. The purpose of this study was to evaluate the utility of histopathology; immunohistochemistry (IHC) for CD3, CD20, and Ki-67; and polymerase chain reaction (PCR) for antigen receptor rearrangement (T-cell clonality) in the differential diagnosis of severe IBD vs intestinal lymphoma. Endoscopic biopsies from 32 dogs with severe IBD or intestinal lymphoma were evaluated. The original diagnosis was based on microscopic examination of hematoxylin and eosin (HE)-stained sections alone followed by a second evaluation using morphology in association with IHC for CD3 and CD20 and a third evaluation using PCR for clonality. Our results show that, in contrast to feline intestinal lymphomas, 6 of 8 canine small intestinal lymphomas were EATL type 1 (large cell) lymphomas. EATL type 2 was uncommon. Regardless, in dogs, intraepithelial lymphocytes were not an important diagnostic feature to differentiate IBD from EATL as confirmed by PCR. EATL type 1 had a significantly higher Ki-67 index than did EATL type 2 or IBD cases. Based on the results of this study, a stepwise diagnostic approach using histology as the first step, followed by immunophenotyping and determining the Ki67 index and finally PCR for clonality, improves the accuracy of distinguishing intestinal lymphoma from IBD in dogs.

Inducción experimental de la enfermedad de Perthes en corderos / Experimental induction of Perthes disease in lambs

Objetivo: Establecer un modelo experimental sencillo, reproducible y seguro para conocer el desarrollo de la necrosis vascular isquémica de la cadera en el cordero. Material y metodología: Utilizamos 15 corderos (10 machos y 5 hembras) de 4 semanas de edad, divididos en un grupo control (7 animales) y otro grupo experimental (8 animales), a los que se provocó la isquemia de la extremidad proximal del fémur. Se efectuaron radiografía convencional y resonancia nuclear magnética. Tras el sacrificio de los animales, a la 4.a, 8.a y 12.a semanas poscirugía, extrajimos y medimos la cabeza femoral. Una vez fijada la pieza obtuvimos cortes histológicos de diferentes zonas que se tiñeron con hematoxilina-eosina. Resultados: Radiográficamente disminuyó la altura y aumentó la anchura de la cabeza femoral, más evidente a partir de la 4.a semana. No objetivamos cambios en la altura del pilar lateral ni en la distancia artículo-trocantérea. El grupo experimental macroscópicamente demostró hipertrofia y aplanamiento progresivo de la cabeza. A las 4 semanas de la cirugía aparecieron zonas de necrosis en el cartílago articular, una médula ósea más densa y menor altura de la fisis. Los vasos estaban engrosados por proliferación de la capa media y de la adventicia. A las 8 semanas encontramos una fibrosis subcondral, con un cartílago articular irregular, adelgazado y desvitalizado, y áreas de angiogénesis con grasa en el hueso subcondral. A las 12 semanas apreciamos el cierre de la fisis, áreas condrales en las trabéculas óseas y células adiposas en la médula diafisaria. Conclusión: Aunque los cambios histológicos son compatibles con necrosis de la cabeza femoral, las pruebas de imagen obtenidas no se asemejan a la enfermedad de Perthes, por lo que desaconsejamos este modelo experimental para el estudio de esta entidad (AU)

Objective: To establish a simple, reproducible and safe experimental model, for the development of ischemic vascular necrosis of the hip in the lamb. Material and methods: We used 15 lambs (10 males and 5 females) aged four weeks, divided into a control group (7 animals) and an experimental group (8 animals) producing ischemia in the proximal femur. Standard radiography and MRI were performed. The animals were euthanised at the 4th, 8th and 12th weeks after surgery. The femoral heads were extracted and measured and a histological analysis was performed with hematoxylin-eosin staining. Results: Decreased height and increased width of the femoral head was observed in the X -rays, particularly after the 4th week. We did not observe any changes in the height of the lateral pillar or trochanteric distance. The experimental group showed macroscopical hypertrophy and progressive flattening of the head. At 4 weeks necrotic areas in articular cartilage were observed, bone marrow was dense and the growth cartilage height was lower. The vessels were thickened by proliferation of the medial and adventitia layers. At 8 weeks, we found fibrosis in the subchondral bone with thinned and devitalized angiogenesis fat areas. The articular cartilage showed irregularities. At 12 weeks the closure of the physis was noted, as well as chondral areas in the trabecular bone and fat cells in the methaphysis. Conclusion: Although the histological changes are consistent with necrosis of the femoral head, the images obtained did not resemble Perthes disease, so we do not advise this experimental model for the study of this disease (AU)

[Experimental induction of Perthes disease in lambs]. / Inducción experimental de la enfermedad de Perthes en corderos.

OBJECTIVE: To establish a simple, reproducible and safe experimental model, for the development of ischemic vascular necrosis of the hip in the lamb. MATERIAL AND METHODS: We used 15 lambs (10 males and 5 females) aged four weeks, divided into a control group (7 animals) and an experimental group (8 animals) producing ischemia in the proximal femur. Standard radiography and MRI were performed. The animals were euthanised at the 4th, 8th and 12th weeks after surgery. The femoral heads were extracted and measured and a histological analysis was performed with hematoxylin-eosin staining. RESULTS: Decreased height and increased width of the femoral head was observed in the X-Rays, particularly after the 4th week. We did not observe any changes in the height of the lateral pillar or trochanteric distance. The experimental group showed macroscopical hypertrophy and progressive flattening of the head. At 4 weeks necrotic areas in articular cartilage were observed, bone marrow was dense and the growth cartilage height was lower. The vessels were thickened by proliferation of the medial and adventitia layers. At 8 weeks, we found fibrosis in the subchondral bone with thinned and devitalized angiogenesis fat areas. The articular cartilage showed irregularities. At 12 weeks the closure of the physis was noted, as well as chondral areas in the trabecular bone and fat cells in the methaphysis. CONCLUSION: Although the histological changes are consistent with necrosis of the femoral head, the images obtained did not resemble Perthes disease, so we do not advise this experimental model for the study of this disease.

Pathology and diagnostic criteria of Clostridium difficile enteric infection in horses.

Clostridium difficile is commonly associated with diarrhea and colitis in humans and other mammals, including horses. To this date, the epidemiologic, microbiologic, clinical, and diagnostic aspects of C. difficile-associated disease (CDAD) in horses have been thoroughly described. However, reports describing the enteric pathology of this disease in horses are limited. This study presents a comprehensive description of the pathologic characteristics of CDAD in 21 horses and discusses the criteria for the diagnosis of the disease. Case selection was based on C. difficile A/B toxins detection (enzyme-linked immunosorbent assay) in intestinal content samples accompanied by compatible gross and microscopic enteric lesions. Grossly, multifocal, segmental, or diffuse hemorrhage; congestion; and/or marked gelatinous edema of the intestinal wall with abundant bloody or green watery contents were observed. Histologically, the most common lesion was severe necrotizing or necrohemorrhagic enteritis, colitis, or typhlocolitis, with mucosal and/or submucosal thrombosis and marked submucosal edema. The pathology of CDAD in horses is similar to that caused by other equine enteric pathogens; therefore, a definitive diagnosis requires detection of C. difficile A/B toxins in the intestinal contents.

Relationships between fertility and some parameters in male broiler breeders (body and testicular weight, histology and immunohistochemistry of testes, spermatogenesis and hormonal levels).

In a farm of grandparent broiler breeder chickens, we followed the development of 350 roosters from 6 to 55 weeks of age. Data collected and evaluated from these males included body weight, testicular weight, histologic and immunohistochemistry studies of the testes, hormone levels (testosterone, estradiol and corticosterone) and sperm production. The objective was to understand the factors that affect or influence hatch loss that is commonly observed after 45 weeks of age in breeder flocksare often correlated to broiler breeder male chickens. The results of this study showed that in conjunction with the weight of the rooster, the testicle weight increases quickly after the rooster receives light stimulation. At an older age, the study showed that there is a process of testicular shrinkage, and the same effect is seen in sperm production and testosterone levels in broiler breeder roosters. From the histology evaluation, we defined 5 histologic phases that illustrate the evolution of the testicular tissue: perinatal, infantile, puberty, adult and senile. We observed that the adult males with a body weight <3800 g were infertile or had subfertile levels and also had low levels of testosterone and high levels of corticosterone. In contrast, the heaviest males showed correct testicular vitality, high levels of testosterone and low levels of corticosterone. However, the roosters that had acquired this high body weight were also at risk of having less complete copulations because of their physical mass. The loss of uniformity of the males and the appearance of hierarchies within the flock accompany a decline in the percentages of hatches as a consequence of the poor confirmation of the males for copulation or the restriction to the access to the females. Results of this study show that the decrease in fertility from 45 weeks of age has been associated with a decline in testicular weight, sperm production and the testosterone levels in animals with a sub-par weight. Likewise, decreasing hatch in older flocks may also result from a loss of conformation, and the lack of complete copulations is possible because of animals that are grossly overweight.

Feline gastrointestinal lymphoma: mucosal architecture, immunophenotype, and molecular clonality.

Gastrointestinal lymphomas were identified in 120 cats between 1995 and 2006. Lymphomas were classified according to the World Health Organization (WHO) scheme. Cats with mucosal T-cell lymphoma (n = 84) predominated and had a median survival of 29 months. Mucosal T-cell lymphoma matched WHO enteropathy-associated T-cell lymphoma (EATCL) type II. Epitheliotropic T-cell infiltrates were present in 62% of cats and occurred as clusters or diffuse infiltrates of small to intermediate-sized T cells in villous and/or crypt epithelium. Similar lymphocytes infiltrated the lamina propria in distinctive patterns. Cats with transmural T-cell lymphoma (n = 19) had a median survival of 1.5 months. Transmural T-cell lymphoma matched WHO EATCL type I. Epitheliotropic T-cell infiltrates were present in 58% of cats. Large lymphocytes (n = 11), mostly with cytoplasmic granules (LGL-granzyme B+) (n = 9) predominated. Transmural extension across the muscularis propria characterized the lesion. Both mucosal and transmural T-cell lymphomas were largely confined to the small intestine, and molecular clonality analysis revealed clonal or oligoclonal rearrangements of T-cell receptor-γ in 90% of cats. Cats with B-cell lymphoma (n = 19) had a median survival of 3.5 months. B-cell lymphomas occurred as transmural lesions in stomach, jejunum, and ileo-cecal-colic junction. The majority were diffuse, large B-cell lymphomas of centroblastic type. In conclusion, T-cell lymphomas characterized by distinctive mucosal architecture, CD3 expression, and clonal expansion predominated in the feline gastrointestinal tract.

Canine gastric carcinoma: immunohistochemical expression of cell cycle proteins (p53, p21, and p16) and heat shock proteins (Hsp27 and Hsp70).

Immunohistochemical staining for cell cycle proteins and heat shock proteins was performed on 17 canine gastric carcinomas. The immunoexpression of p53, p21, p16, Hsp27, and Hsp70 was investigated. A study was conducted to determine the histological type and parameters related to tumor malignancy. Possible associations and trends were assessed between the immunoexpression of each protein and tumor type as well as specific parameters of malignancy. High intratumor frequency of cellular p53 immunostaining was observed (61.96% average), but lower frequencies of p21 and p16 expression were present (34.65% and 10.41%, respectively). The p53 overexpression was associated with tumor infiltration (P = .0258). Expression of p21 was lower in undifferentiated carcinomas, and the loss of expression was associated with histopathological parameters characteristic of a poor prognosis such as lymphatic vessel invasion (P = .0258). The lack of p16 immunoreactivity was related to histopathological characteristics of malignancy such as the presence of evident and multiple nucleoli (P = .0475). In contrast, deep tumor infiltration was observed in those carcinomas with a high p16 index (P = .0475). Hsp70 appeared to be overexpressed in all gastric neoplasms included in this study. This is in contrast to Hsp27, because a group of tumors showed complete lack of Hsp27 immunoexpression, whereas the others displayed extensive Hsp27 immunostaining. The differences in Hsp27 did not correlate with any of the histopathological parameters, but Hsp27 immunoexpression was higher in the undifferentiated carcinoma. No significant differences in the expression of the proteins were found in canine gastric carcinomas according to their histological type. These findings may be useful for establishing a prognosis for canine gastric carcinoma.

Influence of Neospora caninum infection in BALB/c mice during pregnancy in post-natal development.

The influence of Neospora caninum infection during pregnancy on the post-natal period has been poorly investigated. In a previous study, we suggested that infection with N. caninum during pregnancy could affect the normal post-natal development of the offspring. For this reason, in the present work we evaluated the influence of N. caninum infection in pregnant BALB/c mice at days 0, 7 and 14 of gestation (groups A, B and C, respectively) on the post-natal development of the offspring from birth to day 60 post-partum (PP). Morbidity and mortality, vertical transmission, and histopathological lesions were investigated. The humoral immune response (IgG) of pups was also evaluated. Results showed that infection with N. caninum during pregnancy had fatal consequences for pups, especially during mid-gestation (day 7). Infection provoked a delay in the general development of neonates, clinical signs compatible with neosporosis and severe histopathological lesions. A high mortality rate was found in all infected groups. A 69% of mortality rate was found in group A, a 100% in group B and a 46% in group C. Necrotizing encephalitis and multifocal hepatocellular necrosis were the most severe lesions found. All neonates, except four animals from group C, had antibodies against N. caninum but the immune response was not sufficient to control parasite infection. We have demonstrated that extension of the observation period after N. caninum infection permits a more accurate study of vertical transmission, the major route of parasite transmission, and mortality rates. We propose that infection at mid-gestation (day 7) in BALB/c mice and its study during the post-natal period constitutes a valuable experimental model for testing new chemotherapeutic agents and vaccines designed to protect against congenital neosporosis, in order to select effective protocols before its use on bovine.

Evaluation of Neospora caninum and Toxoplasma gondii infections in alpaca (Vicugna pacos) and llama (Lama glama) aborted foetuses from Peru.

The aim of this study was to investigate the participation of Neospora caninum and Toxoplasma gondii in abortion cases of Peruvian llamas and alpacas. Fifteen aborted foetuses were recovered from two main rearing areas of camelids in Peru (Central or South Andean region). Foetal histopathology was used to detect the presence of protozoal-associated lesions in target organs. N. caninum and T. gondii infections were confirmed by immunohistochemistry (IHC) combined with PCR and by PCR alone, respectively. The influence of the species (llama and alpaca), foetal age (first, second and third gestational periods) and geographical location (Central or South Andean region) of the foetuses was also studied. Thirteen of the samples (26%, 13/50) showed lesions suggestive of protozoal infection. N. caninum infection was detected by either IHC or specific PCR in 14 out of 50 foetuses (28%), of which 8 also showed protozoal-associated lesions. T. gondii DNA was not detected in any of the foetuses analysed. Protozoal infection was more frequent in the foetuses from the second gestational period (P<0.05, Fisher F-test). No significant association was observed between protozoal infection and species or geographical location (P>0.05, chi2 test). The results of the present study indicate that neosporosis should be included during the differential diagnosis of abortion in llamas and alpacas.

Molecular characterisation of BSR4, a novel bradyzoite-specific gene from Neospora caninum.

Here we present the identification and cloning of the NcBSR4 gene, the putative Neospora caninum orthologue to the Toxoplasma gondii TgBSR4 gene. To isolate NcBSR4, genome walking PCR was performed on N. caninum genomic DNA using the expressed sequence tag NcEST3c28h02.y1 sequence, which shares a 44% identity with the TgBSR4 gene, as a framework. Nucleotide sequencing of amplified DNA fragments revealed a single uninterrupted 1227 bp open reading frame that encodes a protein of 408 amino acids with 66% similarity to the TgBSR4 antigen. A putative 39-residue signal peptide was found at the NH2-terminus, followed by a hydrophilic region. At the COOH-terminus, a potential site for a glycosylphosphatidylinositol anchor was identified at amino acid 379. A polyclonal serum against recombinant NcBSR4 protein was raised in rabbits, and immunolabelling demonstrated stage-specific expression of the NcBSR4 antigen in N. caninum bradyzoites produced in vitro and in vivo. Furthermore, RT-PCR analysis showed a slight increase of NcBSR4 transcripts in bradyzoites generated during in vitro tachyzoite-to-bradyzoite stage-conversion, suggesting that this gene is specifically expressed at the bradyzoite stage and that its transcription relies on the switch to this stage.

Maxillary ameloblastic carcinoma in an Alaskan Malamute.

A 3-year-old female Alaskan Malamute was submitted to the Veterinary Teaching Hospital of Madrid with a mass in the right maxilla. It was ulcerated, covered by a yellowish infected membrane, and painful. The histologic examination showed a neoplastic proliferation of palisading cells distributed irregularly in cords. The tumor was diagnosed as an ameloblastic carcinoma because of the histologic signs of malignancy, the aggressive behavior, and the absence of distal metastasis. This is the first description of an ameloblastic carcinoma in a dog.

Clinico-pathological findings in cattle exposed to chronic bracken fern toxicity.

AIM: To describe changes in blood and urine analytes in a large group of cattle exposed to chronic bracken fern toxicity, in order to identify parameters of potential diagnostic value. METHODS: The study was conducted on two livestock farms on which bovine enzootic haematuria (BEH) was known to occur; Farm A grazed a local breed of cows and Farm B grazed Friesians. Group A1 comprised 66 cows from Farm A, Group B 54 cows from Farm B, and Group A2 13 heifers from Farm A. Ten healthy cows were used as controls. A complete physical examination was performed (Group A1), and blood (all groups) and urine (Groups A1 and B) samples were collected. Necropsies and histopathology were undertaken on four cows. RESULTS: Anaemia, leucopenia, monocytosis, thrombocytopenia, hypergammaglobulinaemia, microhaematuria and proteinuria were detected. Multivariate statistical analyses established three phases of the disease of increasing severity; an initial phase, characterised by an extremely high monocytosis and otherwise normal parameters; an intermediate phase, characterised by monocytosis and moderate changes to other analytes; and a final phase, characterised by normal levels of monocytes and many changes to other analytes. CONCLUSIONS AND CLINICAL RELEVANCE: Monocytosis, detected in 31% of the younger animals, could represent an initial response to consumption of bracken fern and might be useful as an early haematological marker of BEH.

Comparison of Neospora caninum distribution, parasite loads and lesions between epidemic and endemic bovine abortion cases.

It has been suggested that the abortion herd pattern could influence bovine foetal neosporosis. Here, a comparison of (i) Neospora caninum DNA-detectability by PCR, (ii) N. caninum-associated lesions and (iii) parasite loads in target organs was made between epidemic and endemic abortion cases. We observed that N. caninum DNA was predominantly detected in more than one organ in the foetuses from herds with epizootic rather than endemic abortion cases (P<0.05, Fisher F-test). The highest parasite burdens were found in the heart in foetuses from outbreaks of epidemic abortion and in the brain in endemic cases (P<0.05, Kruskal-Wallis H-test). Moreover, foetuses from epidemic outbreaks had significantly higher parasite burdens in heart (P<0.05, Mann-Whitney U-test) than endemic abortion cases. Epidemic abortion cases showed higher lesion frequencies in liver (P<0.05, Fisher F-test). This report confirms that the abortion herd pattern is an important factor that influences pathogenesis in natural N. caninum infections.

Lingual calcinosis circumscripta in a dog.

Calcinosis circumscripta occurs rarely in dogs and cats, especially in the oral cavity. This case report describes the diagnosis and management of lingual calcinosis circumscripta in an eight-month-old, intact/male German shepherd-cross dog. An excellent prognosis for this disease may be achieved when the patient is managed as described in this report.

Pathological alterations caused by Anoplocephala perfoliata infection in the ileocaecal junction of equids.

The pathological alterations caused by Anoplocephala perfoliata in the ileocaecal junction of 28 equids slaughtered in an abattoir in Madrid (Central Spain) are described. The lesions were scored in grades based on the intensity of the damage and were related to the tapeworm number observed. The first grade (grade I) of alterations consisted of a slight enteritis associated with focal erosions observed in 43% of parasitized animals with low parasitic burden (1-26 tapeworms). The second grade (grade II) was a focal pseudomembranous enteritis, present in the ileocaecal junctions of 36% infected animals with moderate to high burden (23-188 tapeworms), and the third grade (grade III) was a regional necrotizing enteritis, present in the animals (21%) with the highest burden (72-248 tapeworms). The possible role of the lesions caused by this parasite in the aetiology of colic is discussed.