Quantitative risk assessment of introduction of BVDV and BoHV-1 through indirect contacts based on implemented biosecurity measures in dairy farms of Spain.

A stochastic quantitative risk assessment model was developed to estimate the annual probability of introduction of bovine viral diarrhea virus (BVDV) and bovine herpesvirus 1 (BoHV-1) on 127 dairy farms through indirect contacts. Vehicles transporting calves, cattle to slaughterhouse, dead animals, and mixture of feed, as well as visits by veterinarians and hoof trimmers, farm workers and contacts with neighbors were considered in the model. Data from biosecurity questionnaires of each farm, scientific literature and expert opinion from field veterinarians, animal vehicle drivers, hoof trimmers and personnel from rendering transport companies were used to estimate values for input parameters. Results showed that the annual probability of introducing BVDV or BoHV-1 through indirect contacts was very heterogeneous. The overall distribution of median values for each farm ranged from 0.5 to 14.6% and from 1.0 to 24.9% for BVDV and BoHV-1, respectively. The model identified that providing protective clothing and boots to visits, not allowing the animal vehicle driver to come into contact with animals present on the farm and ensuring that calf vehicles arrived empty, were the measures with the highest impact on the probability of infection for most farms. This model could be a useful tool to show the impact of the measures to farmers and veterinarians, thus increasing their awareness on biosecurity. In addition, it could support decision making on which measures should be prioritized in dairy cattle herds to reduce the probability of introduction of diseases.

Short communication: Risk factors associated with Mycobacterium avium ssp. paratuberculosis introduction into dairy herds in Galicia, northwestern Spain.

This study assessed potential risk factors associated with introduction of Mycobacterium avium ssp. paratuberculosis (MAP) into dairy cattle herds in the Galicia region, northwestern Spain. The study was carried out with data collected from 93 dairies enrolled in a voluntary MAP control program. Information on potential risk factors was obtained through personal interviews with the farmers and veterinarians in charge of the control program of each farm. In addition, blood samples were taken annually over 2 years from cows on the farms in the program, and analyzed with a commercial ELISA to detect antibodies to MAP. Fecal samples of all ELISA-positive cows were analyzed using PCR. Based on χ2 test and Fisher's exact test, purchase practices, shared manure truck, shared materials, and visitors per month who contacted animals were found to be significantly associated with farm MAP infection status. Multiple logistic regression indicated that purchase practices and herd size (included as a potential confounder) are the variables that best predict MAP status.

Development of a quantitative risk assessment of bovine viral diarrhea virus and bovine herpesvirus-1 introduction in dairy cattle herds to improve biosecurity.

A quantitative risk assessment model was developed to estimate the annual probability of introducing bovine viral diarrhea virus (BVDV) and bovine herpesvirus 1 (BoHV-1) at the farm level through animal movements. Data from 2017 official animal movements, biosecurity questionnaires, scientific literature, and expert opinion from field veterinarians were taken into consideration for model input parameters. Purchasing or introducing cattle, rearing replacement heifers offsite, showing cattle at competitions, sharing transport vehicles with other herds, and transporting cattle in vehicles that have not been cleaned and disinfected were considered in the model. The annual probability of introducing BVDV or BoHV-1 through infected animals was very heterogeneous between farms. The median likelihoods of BVDV and BoHV-1introduction were 12 and 9%, respectively. Farms that purchased cattle from within their region (i.e., local movements) and shared transport with other farms had a higher probability for BVDV and BoHV-1 introduction. This model can be a useful tool to support decision-making on biosecurity measures that should be prioritized to reduce the probability of introduction of these 2 diseases in dairy herds.

Robot milking and relationship with culling rate in dairy cows.

Cow routines and behavioral responses are altered substantially following the installation of robot milking. The present study was designed to analyze the effect that switching from milking parlor to automatic milking system (AMS) had on the culling rate (due to various causes) of dairy cattle. For this purpose, culling records and causes for culling were tracked in 23 dairy farms in the Galicia region (NW Spain). The animals in these farms were monitored for 5 years. For the present study, that length of time was divided into three different stages, as follows: 2 years before switching from a milking parlor to AMS (stage 1), the 1st year following the implementation of AMS (stage 2) and the 2nd and 3rd years succeeding the implementation of AMS (stage 3). Cox models for survival analysis were used to estimate the time to culling due to different reasons during stage 1 in relation to stages 2 and 3. The data indicated that the risk of loss due to death or emergency slaughter decreased significantly following the installation of AMS. In contrast, the risk of culling due to low production, udder problems, infertility or lameness increased significantly. Low-production cows (such as cows in advanced lactation due to infertility) or sick cows (such as mastitic or lame cows) allegedly have a noticeable effect both on the performance and the amortization of the cost of AMS, which in turn would lead to a higher probability of elimination than in conventional systems.

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.

Study on some risk factors and effects of bovine ketosis on dairy cows from the Galicia region (Spain).

The study was designed to determine the relation between ß-hydroxybutyrate (BHB) concentrations in milk from dairy cows after calving and the length of the previous lactation, the dry period and the 305-day normalized production, and to assess the influence of BHB concentrations on culling and test-day milk productions and somatic cell counts (SCC) throughout the lactation that followed the BHB measurement. The data used in the study were obtained from 59 187 cows in the Galicia region (Spain). BHB determination was performed using Fourier-transformed infrared spectrometry from the milk samples collected from each cow on the first post-partum test day. For statistical analysis, the following methods were applied: (i) ordinal regression to assess the effect of the length of the previous lactation, the dry period and the 305-day normalized milk production on milk BHB, (ii) a Cox model to estimate the influence of the BHB concentration on risk of culling (overall and for a variety of reasons) and (iii) linear regression to assess the link between BHB and the milk yield and SCC obtained from each of the tests day performed throughout lactation. The probability of having higher BHB concentrations increased when the length of the previous lactation (p = 0.006), the dry period (p = 0.003) and the 305-day normalized milk yield (p = 0.005) increased. However, the slight increase observed (especially for the case of the dry period and the 305-day milk yield) would not justify that measures be implemented to reduce these traits. Higher concentrations of BHB led to an increased risk of culling due to 'death' (p ≤ 0.001) and 'urgent slaughter' (p ≤ 0.002) (both causes of involuntary culling). It also led to a reduction in milk production (p < 0.001) and an increase in SCC (p < 0.001) in the post-partum; from that moment onward (including peak lactation), there were no differences in those two parameters depending on the BHB levels.

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.

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.

The use of crossbreeding with beef bulls in dairy herds: effects on calving difficulty and gestation length.

This study was designed to analyse the evolution in the use of beef bull semen for dairy cattle insemination and, mainly, to assess calving difficulty, gestation length and proportion of stillbirths after breeding pure Holsteins or crossbreeding. Data were collected during 2004 to 2011 for 552 571 Holstein calvings (457 070 Holstein × Holstein, 43 384 Holstein × Limousine, 32 174 Holstein × Belgian Blue and 19 943 Holstein × Galician Blonde). The highest calving difficulty, compared with pure Holsteins was for crosses with Belgian Blue followed by Limousine and Galician Blonde. The Holstein × Limousine and Holstein × Galician Blonde crossbred calves had significantly longer gestation lengths than Holstein × Holstein and Holstein × Belgian Blue calves. Between the latter two, pure Holstein had the shortest gestation length. Calving difficulty and gestation length decreased as the age of the dam advanced. The most difficult calvings were observed in twin calvings, followed by the calvings of male calves and female calves. The gestations leading to the birth of male calves were longer than those leading to female calves and twin calves. Stillbirths were not related to the breed used for mating. Through examining these parameters, sire breed should be considered when selecting a beef breed for the insemination of milk-producing dams.

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.

30 años del Grupo Español de Dermatopatología / The Spanish Dermatopathology Group-30 Years On

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Pero ¿hubo alguna vez una clasificación de Clark de los melanomas? / But was there ever a Clark classification of melanomas?

Desde hace 40 años, la inmensa mayoría de los dermatólogos y los patólogos de todo el mundo ha aceptado y ha empleado la clasificación de Clark de los melanomas cutáneos. Sin embargo, tras una cuidadosa relectura de los artículos fundamentales de Clark y sus colaboradores, hemos podido comprobar que tal clasificación ha sido en realidad muy efímera. Tras distinguir en 1968 entre melanoma nodular, melanoma de extensión superficial (MES) y melanoma del lentigo maligno (MLM), la inclusión en 1979 del melanoma lentiginoso acro (MLA) como un nuevo subtipo de melanoma fue la primera avería seria de la clasificación, ya que un melanoma lentiginoso y de aparición tardía (como el MLM) se localizaba, a diferencia de éste, en las zonas menos fotoexpuestas de la piel. Posteriormente, los mismos autores comprobaron que, contrariamente a su idea inicial, el pronóstico del MLM era el mismo que el de los demás subtipos a igualdad de espesor, según Breslow. Finalmente, diversas observaciones de los mismos autores fueron poniendo de manifiesto su creciente dificultad para distinguir al microscopio entre MLM, MES y MLA, salvo que tuviesen en cuenta la localización. Es decir, que hoy por hoy las posibles diferencias morfológicas entre uno y otro caso de melanoma cutáneo no conllevan demostradas diferencias pronósticas, y las diferencias morfológicas que puedan encontrarse se deben más a la diferente localización que a la propia neoplasia (AU)

For the past 40 years, the Clark classification of cutaneous melanoma has been accepted and used by the vast majority of dermatologists and pathologists throughout the world. However, after careful rereading of the most relevant articles by Clark and his collaborators, we can affirm that the classification was only ever of passing validity. After distinguishing between nodular melanoma, superficial spreading melanoma (SSM), and lentigo maligna melanoma (LMM) in 1968, the inclusion of acral-lentiginous melanoma (ALM) in 1979 as a new subtype was the first serious setback for the classification; in contrast to ALM, late-onset lentiginous melanomas, such as LMM, were situated on areas of skin with less exposure to sunlight. Later, the same authors found that, contrary to their initial belief, the prognosis of LMM was the same as that of other subtypes with the same Breslow thickness. Finally, a number of observations by the same authors made ever clearer the increasing difficulty for distinguishing microscopically between LMM, SSM, and ALM, except by taking their localization into consideration. This means that, today, the possible morphological differences between one case of cutaneous melanoma and another are of no proven prognostic implication. In addition, the morphological differences that can be found are much more closely related to the different localization that to the tumor itself (AU)

[But...was there ever a Clark classification of melanomas?]. / Pero...hubo alguna vez una clasificación de Clark de los melanomas?

For the past 40 years, the Clark classification of cutaneous melanoma has been accepted and used by the vast majority of dermatologists and pathologists throughout the world. However,after careful rereading of the most relevant articles by Clark and his collaborators, we can affirm that the classification was only ever of passing validity. Today, the possible morphological differences between one case of cutaneous melanoma and another are of no proven prognostic implication. In addition, the morphological differences that can be found are much more closely related to the different localization that to the tumor itself.

Effect of the sampling device on fat and protein variation in cow milk samples obtained for official milk recording.

The paper was designed to assess the effect of the type of milk meter used by dairy farms for official milk testing to obtain individual milk samples for fat and protein determination. This study was performed in 2006 in Galicia, which is Spain's main dairy cattle region; data were collected from 2,149 Holstein cow farms enrolled for official milk recording. Findings indicate that the milk sampling system used has a substantial effect on the fat contents recorded for individual milk samples. Such variation could arise from poor functioning of the equipment, inadequate mixing of the milk portion from which the sample is obtained, or from the nonproportionality of the sample. Samples obtained from only one point in the milk line may not be representative of all the milk produced. The most significant differences are observed in the percentage of fat, whereas differences in percentage of protein are smaller. The sampling method used should, but does not always, provide a representative sample of the milk from a single milking.

¿Tienen ustedes una definición de nevo melanocítico típico? / Do You Have a Definition for Typical Melanocytic Nevus?

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Application of ATP bioluminescence for evaluation of surface cleanliness of milking equipment.

The ATP bioluminescence method was used to evaluate the cleanliness of milking equipment surfaces (teat cup rubbers, teat dip containers, milk receivers, and pipeline joints) in dairy farms in Galicia (northwest Spain) with parlour, pipeline tie-stall or bucket tie-stall milking systems. The cleanest surfaces were teat cup rubbers. The use of non-chlorinated water for cleaning, and of pipeline or bucket tie-stall milking systems, was associated with high ATP bioluminescence values. However, ATP bioluminescence values only explained 12% of the variability in bulk-tank bacterial count; this is attributable to the importance of other factors (notably the correct functioning of the tank cooling system) for maintenance of low bacterial count.

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.