Use of topical local anesthetics to control pain during treatment of hoof lesions in dairy cows.

Hoof pathologies in dairy cows have a major effect on both production and animal welfare. Trimming of excess or diseased hoof tissue is essential for the treatment of many of these conditions. Trimming hoof lesions can cause severe pain, resulting in adverse behavioral responses with risk for animal and human safety. Interventions are usually carried out by nonveterinary technicians in the absence of pain management training. Pain control during trimming is not only an ethical obligation but also allows for better manipulation and more meticulous treatment. The aim of this study was to test the efficacy of Tri-Solfen (Bayer Australia Ltd., Pymble, NSW, Australia), a combination of local anesthetics in a topical gel form, containing lidocaine, bupivacaine, adrenaline, and cetrimide, for the treatment of pain associated with trimming of hoof lesions. Sixty-two Holstein-Frisian cows were selected for trimming at the drying-off period and were visually scored for lameness before entering the chute. After diagnosis of the hoof lesion but before deep trimming was initiated, each animal was randomly distributed to 2 groups: C, usual trimming with no pain control, and T, trimming with a local anesthetic formulation being applied immediately after live corium was exposed. During curative trimming, behavior observation was conducted by 2 observers blind to treatment. In 27 cows, algometry measurements were performed before and after the procedure to assess animal reaction to pressure. Lameness scoring was again performed as the cow left the chute. Nonparametric tests and ANOVA were performed. Results showed that use of the topical anesthetic formulation significantly reduced reaction to trimming and lameness score after trimming when compared with nontreated animals. Algometry values showed increased pressure threshold after application of topical anesthetics. This study suggests that the use of topical local anesthesia with lidocaine and bupivacaine helps reduce pain associated with corrective trimming of severe hoof lesions, enhancing animal welfare and potentially ensuring safety of trimmers.

Gastrointestinal parasites as a possible threat to an endangered autochthonous Portuguese sheep breed.

Helminth and protozoan infections are responsible for important diseases in grazing sheep, which can be especially threatening in an autochthonous breed at risk of extinction like the Churra Galega Mirandesa Portuguese sheep breed. The aim of the present study was to determine the diversity, prevalence and burden of gastrointestinal parasites in these sheep and to assess the effects of deworming practices, cohabiting animals on the farm and feed management. Coprological qualitative and quantitative analysis (flotation, natural sedimentation and McMaster method) were used to identify and quantify gastrointestinal parasites and a questionnaire was designed and applied. A total of 512 faecal samples were collected from 49 flocks, and 49 replies to the questionnaire were received. Parasites were identified in 100% of the flocks, and in 97% of the samples. The genera or species that have been morphologically identified were: strongyle-type, Nematodirus spp., Skrjabinema spp., Moniezia expansa, Moniezia benedeni, Trichuris spp., Capillaria spp., Eimeria spp., Dicrocoelium spp. and Fasciola hepatica. This is the first report in Portugal of Skrjabinema spp. The burden of parasites' oocysts and eggs per gram in faecal samples ranged, respectively, from 50 to 17,550 for Eimeria spp., and from 50 to 6250 for strongyle-type eggs. Factors affecting parasitic infections were evaluated using a multivariate logistic regression. Grazing time and a lack of anthelmintic treatment were positively associated with Nematodirus spp. infection. This study showed that there is a high prevalence and diversity of gastrointestinal parasites in the Churra Galega Mirandesa sheep breed.

Blood lactate, pH, base excess and pCO2 as prognostic indicators in caesarean-born kids from goats with pregnancy toxaemia.

The objective of this study was to identify the prognostic value for survival of blood parameters in the immediate post-caesarean surgery period in kids born from pregnancy toxaemia (PT) goats. This study involved 10 PT goats, in which a caesarean surgery was performed. Twenty-five kids were born after caesarean surgery of which 16 survived. A blood sample was collected from the jugular vein of the 10 goats and from the kids immediately after caesarean surgery (within 15 min). There were differences between the kids that survived and the kids that did not survive concerning the blood levels of pH (7.22 vs 7.00), base excess (- 9 vs - 18 mmol/L), pCO2 (46 vs 62 mmHg) and L-lactate (5.6 vs 16 mmol/L). Maternal ketoacidosis due to PT has a negative impact on the survival rate of the offspring. This appears to be associated to a metabolic acidosis of the offspring. However, the only blood parameter in which there was a strong association between the maternal and newborn kids was blood urea nitrogen (r = 0.97).

Conservation priorities of Iberoamerican pig breeds and their ancestors based on microsatellite information.

Criollo pig breeds are descendants from pigs brought to the American continent starting with Columbus second trip in 1493. Pigs currently play a key role in social economy and community cultural identity in Latin America. The aim of this study was to establish conservation priorities among a comprehensive group of Criollo pig breeds based on a set of 24 microsatellite markers and using different criteria. Spain and Portugal pig breeds, wild boar populations of different European geographic origins and commercial pig breeds were included in the analysis as potential genetic influences in the development of Criollo pig breeds. Different methods, differing in the weight given to within- and between-breed genetic variability, were used in order to estimate the contribution of each breed to global genetic diversity. As expected, the partial contribution to total heterozygosity gave high priority to Criollo pig breeds, whereas Weitzman procedures prioritized Iberian Peninsula breeds. With the combined within- and between-breed approaches, different conservation priorities were achieved. The Core Set methodologies highly prioritized Criollo pig breeds (Cr. Boliviano, Cr. Pacifico, Cr. Cubano and Cr. Guadalupe). However, weighing the between- and within-breed components with FST and 1-FST, respectively, resulted in higher contributions of Iberian breeds. In spite of the different conservation priorities according to the methodology used, other factors in addition to genetic information also need to be considered in conservation programmes, such as the economic, cultural or historical value of the breeds involved.

Early embryo development, number, quality, and location and the relationship with plasma progesterone in dogs.

This research assessed the developmental stages and morphological quality of dog embryos collected during different stages of pregnancy as well as the relationship with serum progesterone recorded at insemination and embryo collection. Embryos were collected from 23 young mature bitches, that had been inseminated with fresh semen 3-6 days after the LH surge (day 0). Embryo flushing was performed on pregnancy days 8-11 (Group 1), 12-15 (Group 2), or 16-20 (Group 3). The location, number and morphological characteristics of the embryos were evaluated. A total of 120 embryos and 25 unfertilized oocytes were collected from bitches with a total of 156 corpora lutea (CL). The mean total embryo yield (total of embryos/CL) was 76.7 ± 5.9%, and the mean embryo recovery rate (number of flushed embryos/number of CL) was 70.6 ± 6.6%. The mean ovulation rate was 6.8 ± 0.5 and the mean number of embryos per bitch was 5.2 ± 0.6. Oocyte fertilization occurred following oocyte maturation. Most embryos in Group 1 (70.0 ± 18.6%) were collected at the 2 to 16 cell stage. The morula stage was first observed on day 11. Expanded blastocysts (EBLs) and hatched blastocysts were first flushed from the uterus on days 13 and 14, respectively. The EBL was the most abundant stage in Groups 2 and 3. After day 19, some embryos (n = 8) had already adhered to the endometrium. Although most recovered embryos were classified as very good, a greater number of low quality embryos was collected in the later gestational periods. A significant variation in the embryonic stages and location of embryos in early canine pregnancy was observed, as embryos entered the uterus independently of their developmental stage. Embryo yield and quality were independent of the serum progesterone concentration at insemination and recovery.

Q Fever Dairy Herd Status Determination Based on Serological and Molecular Analysis of Bulk Tank Milk.

Ruminants are recognized as the main reservoirs of Coxiella burnetii. EFSA highlighted the lack of knowledge about Q fever prevalence in many European countries. A cross-sectional study was carried out in randomly selected dairy herds (n = 109) from central Portugal to screen for C. burnetii infection and to correlate it with herd factors. Bulk tank milk (BTM) samples from cattle (n = 45) and small ruminant (n = 64) herds were tested by ELISA and PCR. The apparent seroprevalence of Q fever was estimated in 45.9% (95% CI: 36.3-55.7) being higher in small ruminants (51.6; 95% CI: 39.6-63.4) than in cattle (37.8; 95% CI: 25.1-52.4). The shedding of C. burnetii in BTM was detected in 11.9% (95% CI: 7.1-19.4) of BTM, and it was higher in cattle (20%; 95% CI: 10.9-33.8) than in sheep and mixed herds (6.3%; 95% CI: 2.5-15). A high bacterial load (≥ 3 × 10(3) bacteria/ml) was observed in 85% of PCR-positive BTM. A significant correlation was found between the bacterial load and positive samples on ELISA (P < 0.001). Antibody positivity was significantly associated with the increased herd size (P < 0.01) and the occurrence of abortion (P < 0.05), whereas the shedding of C. burnetii was significantly associated with the report of infertility (P < 0.05). The results highlight that serological and molecular methods in combination are a useful tool to screen for Q fever and to clarify the herd infection status. The shedding of C. burnetii through milk is important, especially in dairy cattle, and thus, the role of milk as a potential source of infection among dairy workers should not be neglected. To our knowledge, this is the first study reporting C. burnetii infection in dairy livestock in Portugal showing that Q fever is significant in dairy herds, leading to economic losses and being a risk for public health, which highlights the need of implementation of control measures.

Cryopreservation of in vitro-produced sheep embryos: Effects of different protocols of lipid reduction.

UNLABELLED: The low survival of sheep in vitro-produced (IVP) embryos after cryopreservation is a key limiting step to the widespread of this technology. In the present work, different approaches for enhancing cryosurvival of these embryos were compared: embryo delipidation by centrifugation in the absence or presence of cytochalasin D, a cytoskeleton stabilizer or by embryo culture in the presence of different doses of the trans-10 cis-12-conjugated linoleic acid isomer (CLA). Three experiments were conducted. In experiment 1, IVP blastocysts before vitrification were randomly distributed into four groups: control; centrifuged (cent), cytochalasin D (cyto-D), centrifuged + cytochalasin D (cent + cyto-D). In experiment 2, different doses of CLA (25, 50, and 100 µM) were supplemented during embryo culture before vitrification of blastocysts. A control group ran simultaneously. A third experiment was performed to compare both approaches from the previous ones but without the groups with the worst results (groups: control, cyto-D, cent + cyto-D, CLA25, CLA50). In all experiments, embryos integrity and reexpansion were assessed after warming and after 3 hours of culture. In experiment 1, the postwarming integrity rate was the lowest (P < 0.05) in embryos from the cent group (cent: 50.6 ± 10.3% vs. CONTROL: 74.6 ± 9.2%, cyto-D: 92.3 ± 9.7%, and cent + cyto-D: 90.5 ± 11.2%), whereas the best (P < 0.05) reexpansion scores were obtained in cent + cyto-D embryos (cent + cyto-D: 2.6 ± 0.28 vs. CONTROL: 1.8 ± 0.08, cent: 1.9 ± 0.2, and cyto-D: 1.8 ± 0.31). In experiments 2 and 3, higher (P < 0.05) cleavage rates were observed in CLA25 (50.9 ± 6.2% and 49.2 ± 5.6%, respectively) and CLA50 (48.9 ± 6.2% and 47.6 ± 5.6%, respectively) than those in the control (41.8 ± 6.1% and 40.4 ± 5.4%, respectively) group. In experiment 2, CLA100 presented the lowest (P < 0.002) Day-6 and -7 embryo production rate and quality. After warming, superior (P < 0.02) expansion scores were achieved in CLA25 (3.1 ± 0.29) and CLA50 (3.8 ± 0.17) than in the control (1.9 ± 0.10) group. Similar results were attained in experiment 3. However, although cent + cyto-D embryos showed higher (P = 0.008) postwarming expansion scores than the control (2.8 ± 0.29 vs. 1.9 ± 0.07) group, this score was lower (P = 0.0009) than that in CLA50 embryos (3.8 ± 0.17). In conclusion, our results showed that different protocols of lipid reduction can be successfully applied to improve the cryotolerance of IVP sheep embryos.

Serological evidence of exposure to Coxiella burnetii in sheep and goats in central Portugal.

The recent outbreak of Q fever in The Netherlands warned European health authorities of the need of studying Coxiella burnetii. In Portugal, little is known about C. burnetii infection in animals. A cross-sectional study was designed to investigate the exposure to C. burnetii in sheep and goats in the Central region of Portugal, estimating the herd and individual prevalence. A serosurvey was conducted in a two levels random sampling of 89 herds and 460 animals. Individual blood samples were collected from animals older than 6 months, and specific antibodies anti-C. burnetii were detected by ELISA testing. Results showed a global herd prevalence of 32.6% (95% CI: 23.1-42.1%). Herd prevalence was higher in mixed herds (38.5%; 95% CI: 12-65%) and in sheep herds (37.5%; 95% CI: 21-54%) than in goat herds (28.8%; 95% CI: 17-41%). Global individual prevalence was estimated at 9.6% (95% CI: 6.9-12.2%), and it was higher in goats (10.4%; 95% CI: 7.8-13%) than in sheep (8.6%; 95% CI: 5.8-11.4%). Sample positive percentages (S/P) ranged from 41.5% to 185.9%. S/P percent higher than 100 was found in 18.2% (8/44) of sera from distinct herds. Positive results were significantly associated with goats, older animals and larger herds. These results revealed the presence of C. burnetii in small ruminants evidencing their potential role in the infection cycle.

Fat area and lipid droplet morphology of porcine oocytes during in vitro maturation with trans-10, cis-12 conjugated linoleic acid and forskolin.

Lipid droplets (LD) in porcine oocytes form a dark mass reaching almost all cytoplasm. Herein we investigated changes in fat areas, cytoplasmic tone and LD morphology during in vitro maturation (IVM) of porcine oocytes cultured with 100 µM trans-10, cis-12 conjugated linoleic acid (t10,c12 CLA) or 10 µM forskolin at different time periods. Four groups were constituted: control, excipient, t10,c12 CLA and forskolin, with drugs being supplemented during 44 to 48 h and the initial 22 to 24 h in Experiments 1 and 2, respectively. In Experiment 3, forskolin was supplemented for the first 2 h. Matured oocytes were inseminated with frozen-thawed boar semen and cleavage rate recorded. Before and during IVM, samples of oocytes were evaluated for LD, total and fat areas and fat gray value or for meiotic progression. Results showed that forskolin supplementation during 44 to 48 h or 22 to 24 h inhibits oocyte maturation (exp. 1: forskolin = 5.1 ± 8.0%, control = 72.6 ± 5.0%; exp. 2: forskolin = 24.3 ± 7.4%, control = 71.6 ± 5.6%) and cleavage (exp. 1: forskolin = 0.0 ± 0.0%, control = 55.4 ± 4.1%; exp. 2: forskolin = 8.3 ± 3.3%, control = 54.5 ± 3.0%). Forskolin also reduced oocyte and fat areas. In Experiment 3, forskolin negative effect on oocyte maturation and cleavage disappeared, although minor (P ⩽ 0.03) LD and oocyte fat areas were identified at 22 to 24 h of IVM. Oocytes supplemented with t10,c12 CLA during 44 to 48 h presented a lighter (P ⩽ 0.04) colour tone cytoplasm than those of control and forskolin. In conclusion, t10,c12 CLA and forskolin were capable of modifying the distribution and morphology of cytoplasmic LD during porcine oocyte maturation, thus reducing its lipid content in a time-dependent manner.

Effect of hot-iron disbudding on behaviour and plasma cortisol of calves sedated with xylazine.

We investigated cortisol and behaviour for the first hour after hot-iron disbudding of calves aged 37+/-4 days: disbudded after i/m xylazine (n=10); disbudded after i/m xylazine and regional anaesthesia with lidocaine (n=10); sham-disbudded after xylazine and lidocaine (n=11); sham-disbudded after i/m saline and lidocaine (n=10). Xylazine-treated groups had higher cortisol than saline-treated animals and showed no differences among them at any time. Sham-disbudded calves with xylazine had lower cortisol at 60 min compared with all other times. Xylazine-alone disbudded calves struggled more during the procedure than all other groups. Xylazine-alone disbudded calves showed more ear-flicks at 10, 25 and 40 min and head-shakes at 40 min than all other groups. We conclude that cortisol should not be used as an indicator of pain in disbudded calves while under the sedative effect of xylazine and that some behaviours during and after the procedure are useful in showing that xylazine alone does not control hot-iron disbudding pain.

Indicators of genetic erosion in an endangered population: the Alentejana cattle breed in Portugal.

A study was conducted to characterize genetic diversity in the Alentejana breed of cattle based on its demographic trends and to investigate the major factors affecting genetic erosion in this breed. Herdbook information collected between 1940 and 2004, including pedigree records on 100,562 animals in 155 herds, was used to estimate demographic parameters. The mean generation intervals were 6.0 +/- 2.4 yr and 6.8 +/- 3.2 yr for sires and dams of calves, respectively. Average inbreeding increased steadily over the period analyzed, with an annual rate of inbreeding of 0.33 +/- 0.004% (P < 0.01) and an effective population size of 23.3. In the reference population (28,531 calves born between 2000 and 2003) the average inbreeding was 8.35 +/- 9.02% and nearly 80% of the calves were inbred, whereas the average relationship among all animals was 0.026 +/- 0.040. Nevertheless, the mean relationship was 0.328 +/- 0.264 and 0.022 +/- 0.026 for animals born in the same and in different herds, respectively. The computed genetic contributions to the reference population resulted in estimates for the effective number of founders, ancestors, founding herds, and herds supplying sires of 121.6, 55.0, 17.1, and 26.9, respectively, the 2 most influential herds and ancestors contributing 24.2 and 15.1%, respectively, of the current genetic pool. Of the 671 founding sires, only 24 Y-chromosomes are currently represented, but 1 sire alone contributes nearly 60% of this representation, such that the effective number of Y-chromosomes is only 2.73. The observed inbreeding per herd was, on average, 0.053 +/- 0.071 lower than expected from the relationship among the generation of parents of calves in the reference population, indicating that producers have followed breeding strategies that have kept inbreeding at lower levels than anticipated with random selection and mating. When compared with other cattle breeds, Alentejana has some of the highest levels of mean inbreeding and annual rate of inbreeding, and an effective population size that is nearly half of the minimum recommended for maintenance of genetic variability. These critical indicators demonstrate the need to adopt strategies aimed at minimizing inbreeding to avoid further losses of genetic diversity.

Genetic diversity in native and commercial breeds of pigs in Portugal assessed by microsatellites.

Population structure and genetic diversity in the Portuguese native breeds of pigs Alentejano (AL), Bísaro (BI), and Malhado de Alcobaça (MA) and the exotic breeds Duroc (DU), Landrace (LR), Large White (LW), and Pietrain were analyzed by typing 22 microsatellite markers in 249 individuals. In general, the markers used were greatly polymorphic, with mean total and effective number of alleles per locus of 10.68 and 4.33, respectively, and an expected heterozygosity of 0.667 across loci. The effective number of alleles per locus and expected heterozygosity were greatest in BI, LR, and AL, and least in DU. Private alleles were found in 9 of the 22 markers analyzed, mostly in AL, but also in the other breeds, with the exception of LW. The proportion of loci not in Hardy-Weinberg equilibrium in each breed analyzed ranged between 0.23 (AL) and 0.41 (BI, LW, and Pietrain), mostly because of a less than expected number of heterozygotes in those loci. With the exception of MA, all breeds showed a significant deficit in heterozygosity (F(IS); P < 0.05), which was more pronounced in BI (F(IS) = 0.175) and AL (F(IS) = 0.139), suggesting that inbreeding is a major concern, especially in these breeds that have gone through a genetic bottleneck in the recent past. The analysis of relationships among breeds, assessed by different methods, indicates that DU and AL are the more distanced breeds relative to the others, with the closest relationship being observed between LR and MA. The degree of differentiation between subpopulations (F(ST)) indicates that 0.184 of the total genetic variability can be attributed to differences among breeds. The analysis of individual distances based on allele sharing indicates that animals of the same breed generally cluster together, but subdivision is observed in the BI and LR breeds. Furthermore, the analysis of population structure indicates there is very little admixture among breeds, with each one being identified with a single ancestral population. The results of this study confirm that native breeds of pigs represent a very interesting reservoir of allelic diversity, even though the current levels of inbreeding raise concerns. Therefore, appropriate conservation efforts should be undertaken, such as adopting strategies aimed at minimizing inbreeding, to avoid further losses of genetic diversity.