Short communication: Correlation between within-herd antibody-prevalence and bulk tank milk antibody levels to Mycobacterium avium ssp. paratuberculosis using 2 commercial immunoassays.

The objective of this study was to determine the correlation between the results obtained with the ELISA technique for antibodies to Mycobacterium avium ssp. paratuberculosis in serum and bulk tank milk at the herd level. For this purpose, 203 samples of bulk tank milk were analyzed with 2 commercial ELISA from dairy herds with a prevalence of seropositive animals that was also determined. In regard to the reference test (results in blood serum), the sensitivity of the bulk tank milk test to detect high-positive herds (≥10% seroprevalence) ranged from 85.7 to 71.4%. The specificity to detect herds with no seropositive animals ranged from 70.5 to 53%. In a quantitative approach, Pearson correlation coefficients, reported as a measure of the linear association between herd seroprevalences and transformed optical density values recorded in bulk tank milk, were 0.39 and 0.54 for the studied ELISA. Although the test results were relatively fairly correlated with the within-herd prevalence, the practical utility of bulk tank milk testing for Mycobacterium avium ssp. paratuberculosis seems limited, especially regarding specificity.

Bovine herpesvirus 1: within-herd seroprevalence and antibody levels in bulk-tank milk.

The aim of the present study was to establish a relationship between the results obtained with the enzyme-linked immunosorbent assay (ELISA) technique for antibodies (against bovine herpesvirus 1) in serum and those in milk at the herd level. For this purpose, 275 samples of bulk-tank milk were analysed with glycoprotein E (gE) antibody ELISA and 207 more were analysed with glycoprotein B (gB) antibody ELISA (482 in total). All of these samples came from dairy herds whose seroprevalence was also evaluated. The results of this study were then used to analyse the sensitivity of the bulk-tankmilk test in detecting herds with a high risk of active infection (>60% seroprevalence) and its specificity in detecting those with few (<20%) or no seropositive animals. In regard to the reference test (results in blood serum), the sensitivity of the bulk-tankmilk test in detecting herds with >60% seropositive animals was 100% for both gE and gB ELISAs. The specificity figures, for gE and gB ELISAs, respectively, were 88.4% and 99.1% for infection-free herds and 72.6% and 96% for herds with <20% seroprevalence. In a quantitative approach, Pearson's correlation coefficients, reported as a measure of linear association between herd seroprevalences and transformed optical density values recorded in bulk-tank milk, were -0.63 for gE ELISA and 0.67 for gB ELISA.

Les auteurs présentent une étude visant à faire ressortir la corrélation entre les résultats obtenus à l'échelle du troupeau au moyen d'une épreuve immunoenzymatique (ELISA) pour la détection d'anticorps dirigés contre l'herpèsvirus bovin de type 1 dans des échantillons de sérum et ceux obtenus dans le lait. À cet effet, 275 échantillons de lait de citerne ont été soumis à un test ELISA visant à déceler la présence d'anticorps dirigés contre la glycoprotéine E (gE) du virus, et 207 autres ont été analysés au moyen d'un test ELISA visant à déceler la présence d'anticorps dirigés contre la glycoprotéine B (gB) (482 échantillons analysés au total). La totalité des échantillons provenait d'élevages laitiers dans lesquels la séroprévalence a également été evaluée. Les résultats de l'étude ont ensuite permis d'analyser la sensibilité du test sur le lait de citerne, c'est-àdire la capacité de ce test à détecter les troupeaux présentant un risque élevé d'infection active (séroprévalence > 60 %), ainsi que sa spécificité, c'est-à-dire sa capacité à détecter les troupeaux dans lesquels le pourcentage d'animaux séropositifs était faible (moins de 20 %) ou nul (0 %). Comparativement au test de référence (analyse des échantillons de sérum), la sensibilité des tests ELISA sur le lait de citerne était de 100 % (détection de tous les troupeaux dotés d'au moins 60 % d'animaux possédant des anticorps dirigés contre la glycoprotéine E ou B). En termes de spécificité des tests ELISA anti-gE et anti-gB, les valeurs étaient, respectivement, de 88,4 % et 99,1 % dans les troupeaux indemnes et de 72,6 % et 96 % dans les troupeaux accusant une séroprévalence inférieure à 20 %. Les coefficients de corrélation de Pearson obtenus par une méthode quantitative pour exprimer la relation linéaire entre les prévalences sérologiques et les valeurs de densité optique modifiées dans le lait de citerne étaient respectivement de ­0,63 pour l'ELISA gE et de 0,67 pour l'ELISA gB.

Los autores describen un estudio encaminado a determinar si existe una relación, y de ser así cuál, entre los resultados del ensayo inmunoenzimático (ELISA) de detección de anticuerpos (contra el herpesvirus bovino 1) en suero y los resultados obtenidos al analizar la leche de rebaños enteros. Para ello se sometieron 275 muestras de leche de tanque a la prueba ELISA de detección de anticuerpos contra la glicoproteína E (gE) y otras 207 muestras a la prueba ELISA de detección de anticuerpos contra la glicoproteína B (gB) (esto es, un total de 482 muestras). Todas esas muestras procedían de rebaños lecheros cuya prevalencia serológica también se calculó. A partir de los resultados del estudio se determinó la sensibilidad de la prueba practicada en la leche de tanque para detectar rebaños con un elevado riesgo de infección activa (más del 60% de animales seropositivos) y su especificidad para detectar aquellos rebaños con pocos (menos del 20%) animales seropositivos o ninguno (0%). En comparación con la prueba de referencia (resultados del análisis sérico), la sensibilidad del análisis de la leche de tanque para detectar rebaños con más de un 60% de animales seropositivos fue del 100% en el caso de ambas pruebas ELISA (gE y gB). En cuanto a la especificidad, las técnicas ELISA para la gE y la gB permitieron detectar respectivamente un 88,4% y un 99,1% de los rebaños libres de infección y un 72,6% y un 96% de los rebaños con menos de un 20% de animales seropositivos. El análisis cuantitativo de los resultados deparó coeficientes de correlación de Pearson, utilizados como medida de la relación lineal entre las seroprevalencias de rebaño y los valores transformados de densidad óptica obtenidos en la leche de tanque, de ­0,63 para el ELISA gE y de 0,67 para el ELISA gB.

Monitoring the bulk milk antibody response to bovine viral diarrhea in dairy herds vaccinated with inactivated vaccines.

This study was designed to determine long-term responses in dairy herds after vaccination with 1 of 3 inactivated bovine viral diarrhea virus (BVDV) vaccines with regard to antibodies against p80 protein in bulk tank milk samples, as detected by ELISA. In the present study, 29 dairy herds were vaccinated with Bovilis BVD (MSD Animal Health, Milton Keynes, UK), 11 with Hiprabovis Balance (Laboratorios Hipra, Amer, Spain), and 9 with Pregsure BVD (Zoetis, Florham Park, NJ). In these herds, bulk tank milk samples were collected and examined at the time of the first vaccination and every 6 mo during a 3-yr period. Samples were analyzed with a commercial ELISA test for the p80 protein of BVDV. The results demonstrated that vaccination affected the level of antibodies against p80. Hence, vaccination status should be taken into consideration when interpreting bulk tank milk antibody tests.

Changing expression of chloride channels during preimplantation mouse development.

Plasma membrane chloride channels (ClCs) play important roles in a broad range of cellular processes including cell volume regulation, proliferation, and transepithelial transport, all of which are critical during preimplantation embryonic development. In this study, the molecular and functional expression of voltage-gated ClCs was analyzed throughout preimplantation development of the mouse conceptus. mRNA transcripts for all Clcn genes were detected. Only Clcn1 mRNA showed differential expression in the blastocyst, being detected in the trophectoderm but not in the inner cell mass. CLCN3 protein was detected at low levels in the cytoplasm and plasma membrane in 4-cell embryos and was localized to the apical plasma membrane of the trophoblasts in the blastocyst. Whole-cell patch-clamp recordings demonstrated the presence of a DIDS-sensitive, outwardly rectifying Cl(-) current throughout development, with this conductance being large at the 1-cell, morula and blastocyst stages. A second DIDS-insensitive Cl(-) current, which was inactivated by membrane depolarization, was present in cells differentiating into the trophoblast lineage and during blastocyst expansion. Inhibition of the DIDS-sensitive current and the DIDS-insensitive current, with 9-AC, prevented blastocyst expansion.

Long-term acute infections during a bovine viral diarrhea virus (BVDV) outbreak in dairy farm from Galicia (NW Spain).

An observational study describes an outbreak of bovine viral diarrhea virus (BVDV) in a dairy herd in Spain. The herd was subjected to a voluntary control program. In a sampling carried out in June 2020, bulk tank milk antibody levels increased compared to the previous sampling. Additionally, serum samples from 4 young heifers also tested positive for antibodies. Since the results were consistent with a recent infection, we proceeded to detect possible persistently infected (PI) animals using antigen ELISA (on serum/ear-notch samples), following the program guidelines. From this moment on, 42 animals tested positive for BVDV antigen, of which 17 were under typical acute infection (AI), 13 were deemed as PI, and eight died early on the farm before having information to determine their status. The remaining 4 showed intriguing test results consistent with a long-term AI since they tested BVDV positive in at least two antigen tests more than 3 weeks apart. Thus, one animal was positive until 80 days of age in serum, and others even for longer periods in ear-notch samples, until they finally tested negative for BVDV. Based on these results, longer follow-up may be necessary in BVDV positive animals to accurately confirm persistent infection.

Phase II study of capecitabine as palliative treatment for patients with recurrent and metastatic squamous head and neck cancer after previous platinum-based treatment.

BACKGROUND: Platinum-based therapy (PBT) is the standard therapy for recurrent and/or metastatic head and neck cancer (HNC), but the incidence of recurrence remains high. This study evaluates the efficacy and tolerability of capecitabine as palliative monotherapy for recurrent HNC previously treated with PBT. METHODS: Patients aged 18-75 years, with Eastern Cooperative Oncology Group performance status 0-2, squamous HNC with locoregional and/or metastatic recurrence previously treated with PBT and adequate organ functions, were included. Capecitabine (1.250 mg m(-2) BID) was administered on days 1-14 every 21 days for at least two cycles. RESULTS: A total of 40 male patients with a median age of 58 years were analysed. All patients received a median number of four cycles of capecitabine (range: 1-9) and the median relative dose intensity was 91%. Seven patients were not evaluable for response. Overall response rate was 24.2%. Median time to progression and overall survival were 4.8 and 7.3 months, respectively. Haematological adverse events (AEs) grade 3/4 were reported in six patients. Most common grade 3/4 non-haematological AEs were asthenia (12.5%), palmar-plantar eritrodisestesia (10%), mucositis (10%), dysphagia (10%) and diarrhoea (7.5%). CONCLUSIONS: Capecitabine seems to be an active, feasible and well-tolerated mode of palliative treatment for advanced HNC patients who have previously received PBT schedules.

Management practices associated with Mycobacterium avium subspecies paratuberculosis infection and the effects of the infection on dairy herds.

During 2004, a survey of Mycobacterium avium subspecies paratuberculosis (map) was conducted in 101 randomly selected dairy herds to investigate associations between the infection status of the herds, different management practices, and possible disease indicators, such as indices of mastitis and reproductive performance. Data were collected by means of a questionnaire through personal interviews with the farmers and veterinarians in charge of each farm. At the same time, blood samples were taken from cattle over one year old and analysed with a commercial elisa to detect antibodies to map. Statistical analyses indicated that the following management practices constituted major risk factors: utilisation of colostrum from cows with a previous positive map diagnosis, and housing replacement calves with adult cattle before they were six months old. Seropositivity to map was related to the herds' bulk tank somatic cell counts and incidence of clinical mastitis, but not to their reproductive performance.

Genetic diversity of bovine viral diarrhea viruses from the Galicia region of Spain.

This study examined the frequency and diversity of bovine viral diarrhoea viruses (BVDVs) infecting cattle in Galicia (northwestern Spain). A total of 86 BVDV strains were typed in samples of serum from 79 persistently infected animals and 3 viraemic animals and of abomasal fluid from 4 fetuses. Samples came from 73 farms participating in a voluntary BVDV control programme. Typing was based on a 288-bp sequence from the 5' untranslated region amplified using primers 324 and 326. Of the 86 strains, 85 (98.8 per cent) belonged to species BVDV-1 and 1 (1.2 per cent) belonged to BVDV-2; 73 strains (84.9 per cent) were typed as BVDV-1b, 2 as BVDV-1e and 6 as BVDV-1d. One strain each was typed as belonging to 1a, 1h, 1k and 1l. The sole BVDV-2 strain was classified as 2a. These results identify BVDV-1b as the predominant species, and they indicate the presence of viral types not previously described anywhere in Spain. This is also the first report of BVDV-2 in Galicia and only the second report of BVDV-2 in Spain.

[Whole body PET/CT imaging for detection of metastatic choroidal melanoma]. / PET/TC de cuerpo completo para la detección de metástasis de melanoma coroideo.

Choroidal melanoma is the most common primary intraocular cancer in adults. Metastases are most commonly found in the liver and are rapidly fatal despite aggressive therapy. These metastases have been reported in the follow-up to 50 % of uveal melanoma. However, recent reports have shown that liver ultrasonography and liver function tests have low sensitivity in the diagnosis of metastatic uveal melanoma. The study reports on a patient with a history of choroidal melanoma. Preoperative medical evaluation, including CT imaging of the abdomen, complete blood count and liver enzymes proved negative. A PET/CT was requested and showed lesions in the liver and lungs. Whole-body PET/CT revealed melanoma metastases. The PET/CT is a sensitive tool for the detection and localization of hepatic and extrahepatic metastatic choroidal melanoma.

Neospora caninum seroprevalence in dairy and beef cattle from the northwest region of Spain, Galicia.

Herd and individual animal seroprevalence for Neospora caninum (N. caninum) in dairy, beef and mixed cattle were obtained in all populations within the Galician Farmer Sanitary Defence Associations (ADSG) in 2004. All animals ≥1 year of age were examined serologically by indirect ELISA. 1147 dairy herds (37,090 animals), 1464 beef herds (20,206 animals) and 141 mixed herds (2292 animals) were surveyed. True herd seroprevalence was estimated to be 80.6% (87.7% dairy, 76.7% beef and 78.4% mixed herds), true animal seroprevalence was estimated to be 23.2% (21.9% dairy, 25.1% beef and 24.9% animal to mixed herds), and within-herd seroprevalence was estimated to be 25.4% (23.6% dairy, 28.3% beef and 28.6% to mixed herds). Seropositivity was significantly associated with herd type (higher in dairies), herd size (increased when herd size increases), animal type (higher in beef) and age (lineal increase with the age). Results obtained in this study will be used for the development of a N. caninum control programme in the ADSG in Galicia.

Neospora caninum infection in stray and farm dogs: seroepidemiological study and oocyst shedding.

In a previous study, farm and stray dogs were considered potential high risk populations of Neospora caninum infection in Spain. Consequently, we decided to investigate the significance of N. caninum infection in these populations. Specific antibodies were detected in 120 out of 275 dog sera (43.6%), with titres ranging from 1:50 to 1:800. Differences in seroprevalence between farm (47.5%, 67/141) and stray (39.5%, 53/134) dogs were not significant (P>0.05; χ(2) test), but farm dogs showed significantly higher titres (P<0.01; Student's t-test). N. caninum seroprevalence in farm dogs was associated with increasing age (P<0.01; χ(2) test) and dogs with free access to the farm were more likely to be seropositive than controlled-dogs (P<0.05; χ(2) test). The presence of anti-N. caninum antibodies was more often detected in dogs from farms with 5-20% N. caninum within-herd seroprevalence (56.9%, 37/65) than those from farms with 0-5% seroprevalence (38%, 23/60) (P<0.05; χ(2) test). We microscopically observed N. caninum-like oocysts in the faeces from one farm dog, but the number of oocysts was very low, and the aetiology could not be confirmed. Also, parasite isolation was attempted from fresh neural tissue from stray dogs but was unsuccessful.

PET/TC de cuerpo completo para la detección de metástasis de melanoma coroideo / Whole body PET/CT imaging for detection of metastatic choroidal melanoma

El melanoma coroideo es el tumor maligno ocular primario más frecuente en los adultos y las metástasis más frecuentes son a nivel hepático con un mal pronóstico a pesar de los tratamientos. Estas metástasis se han descrito hasta en el 50 % de los melanomas coroideos. Estudios recientes muestran que la ecografía hepática y las analíticas hepáticas tienen baja sensibilidad en el diagnóstico de las metástasis hepáticas de los melanomas de úvea. Exponemos el caso de un paciente con historia de melanoma coroideo. La exploración preoperatoria, incluyendo TC de abdomen, analítica sanguínea y enzimas hepáticos resultó normal. En un PET/TC realizado mostró lesiones en el hígado y pulmones. El PET/TC de cuerpo entero mostró metástasis de un melanoma uveal. El PET/TC es una herramienta útil para la detección de las metástasis hepáticas y extrahepáticas (AU)

Choroidal melanoma is the most common primary intraocular cancer in adults. Metastases are most commonly found in the liver and are rapidly fatal despite aggressive therapy. These metastases have been reported in the follow-up to 50 % of uveal melanoma. However, recent reports have shown that liver ultrasonography and liver function tests have low sensitivity in the diagnosis of metastatic uveal melanoma. The study reports on a patient with a history of choroidal melanoma. Preoperative medical evaluation, including CT imaging of the abdomen, complete blood count and liver enzymes proved negative. A PET/CT was requested and showed lesions in the liver and lungs. Whole-body PET/CT revealed melanoma metastases. The PET/CT is a sensitive tool for the detection and localization of hepatic and extrahepatic metastatic choroidal melanoma (AU)

[Estimation of infection by Aspergillus fumigatus in mice infected experimentally by the respiratory route]. / Estimación de la infección por Aspergillus fumigatus en ratones infectados experimentalmente por vía respiratoria.

BACKGROUND: To evaluate a new chitin detection technique for the quantification of Aspergillus fumigatus infection in organs from air-borne infected mice. METHODS: Different groups of mice were air-borne infected with A. fumigatus spores. The mortality rate was studied as well as the extension of the infection by means of colony counts (UFC) and quantification of chitin . RESULTS: The inhalation of A. fumigatus spores by non-immunosuppressed mice produces a 34% mortality rate, with an initial invasion of lung tissue followed by the spleen, kidneys, heart and liver thereafter. CONCLUSION: The quantification of chitin in involved organs allowed us to estimate the degree of organ-specific fungal infections and perhaps could be an useful method for studying the pathogenesis of A. fumigatus infections.

Skull metastasis from rectal gastrointestinal stromal tumours

Gastrointestinal stromal tumours (GIST) are the most common mesenchymal neoplasm of the gastrointestinal tract. Rectum localisation is infrequent for these neoplasms, accounting for about 5% of all cases. Distant metastases of GIST are also rare. We present a patient with special features: the tumour is localised in rectum and it has an uncommon metastatic site, the skull, implying a complex differential diagnosis approach (AU)