Genetic diversity, structure, and breed relationships in Iberian cattle

In Iberia there are 51 officially recognized cattle breeds of which 15 are found in Portugal and 38 in Spain. We present here a comprehensive analysis of the genetic diversity and structure of Iberian cattle. Forty of these breeds were genotyped with 19 highly polymorphic microsatellite markers. Asturiana de los Valles displayed the greatest allelic diversity and Mallorquina the least. Unbiased heterozygosity values ranged from 0.596 to 0.787. The network based on Reynolds distances was star-shaped with few pairs of interrelated breeds and a clear cluster of 4 breeds (Alistana/Arouquesa/Marinhoa/Mirandesa). The analysis of the genetic structure of Iberian cattle indicated that the most probable number of population clusters included in the study would be 36. Distance results were supported by the STRUCTURE software indicating a relatively recent origin or possible crossbreeding or both between pairs or small groups of breeds. Five clusters included 2 different breeds (Betizu/Pirenaica, Morucha/Avile?a, Parda de Monta?a/Bruna de los Pirineos, Barros?/Cachena, and Toro de Lidia/Brava de Lide), 3 breeds (Berrenda en Negro, Negra Andaluza, and Mertolenga) were divided in 2 independent clusters each, and 2 breeds were considered admixed (Asturiana de los Valles and Berrenda en Colorado). Individual assignation to breeds was not possible in the 2 admixed breeds and the pair Parda de Monta?a/Bruna de los Pirineos. The relationship between Iberian cattle reflects their geographical origin rather than their morphotypes. Exceptions to this geographic clustering are most probably a consequence of crossbreeding with foreign breeds. The relative genetic isolation within their geographical origin, the consequent genetic drift, the adaptation to specific environment and production systems, and the influence of African and European cattle have contributed to the current genetic status of Iberian cattle, which are grouped according to their geographical origin. The greater degree of admixture observed in some breeds should be taken into account before using molecular markers for genetic assignment of individuals to breeds.     

Role of wild ruminants in the epidemiology of bluetongue virus serotypes 1, 4 and 8 in Spain

ABSTRACT: Although the importance of wild ruminants as potential reservoirs of bluetongue virus (BTV) has been suggested, the role played by these species in the epidemiology of BT in Europe is still unclear. We carried out a serologic and virologic survey to assess the role of wild ruminants in the transmission and maintenance of BTV in Andalusia (southern Spain) between 2006 and 2010.A total of 473 out of 1339 (35.3%) wild ruminants analyzed showed antibodies against BTV by both ELISA and serum neutralization test (SNT). The presence of neutralizing antibodies to BTV-1 and BTV-4 were detected in the four species analyzed (red deer, roe deer, fallow deer and mouflon), while seropositivity against BTV-8 was found in red deer, fallow deer and mouflon but not in roe deer. Statistically significant differences were found among species, ages and sampling regions. BTV RNA was detected in twenty-one out of 1013 wild ruminants (2.1%) tested. BTV-1 and BTV-4 RNA were confirmed in red deer and mouflon by specific rRT-PCR.BTV-1 and BTV-4 seropositive and RNA positive wild ruminants, including juveniles and sub-adults, were detected years after the last outbreak was reported in livestock. In addition, between the 2008/2009 and the 2010/2011 hunting seasons, the seroprevalence against BTV-1, BTV-4 and BTV-8 increased in the majority of provinces, and these serotypes were detected in many areas where BTV outbreaks were not reported in domestic ruminants. The results indicate that wild ruminants seem to be implicated in the dissemination and persistence of BTV in Spain.     

Leishmania spp. in Didelphis albiventris and Micoureus paraguayanus (Didelphimorphia: Didelphidae) of Brazil

Leishmaniasis is kept in nature by the participation of several animal species. This study evaluated the presence of Leishmania spp. in skin samples of free-ranging marsupials Micoureus paraguayanus (n=95) and Didelphis albiventris (n=191), captured in Morro do Diabo State Park and in sections of its surrounding forest, in the region of Pontal do Paranapanema, S?o Paulo State, Brazil. The samples were tested for the presence of kDNA of Leishmania spp. by polymerase chain reaction (PCR) and by real time PCR (qPCR). All samples from D. albiventris tested by PCR were negative for the presence of kDNA of Leishmania spp. However, when tested by qPCR, the positivity was 1.6%. A positivity of 7.4% by PCR and 11.6% by qPCR was observed for M. paraguayanus. Sixty-four per cent (9/14) of positive animals were limited to the same forest fragment. Presence of Leishmania (Leishmania) amazonensis and Leishmania (Viannia) braziliensis was detected in M. paraguayanus samples. While D. albiventris is the most studied marsupial species due to its urban habits, other marsupial species such as M. paraguayanus can be potential reservoirs of Leishmania spp. and should also be studied.     

Spatial and temporal investigation of Echinococcus granulosus coproantigen prevalence in farm dogs in South Powys, Wales

The prevalence of canine echinococcosis appears to have been increasing in mid/south Wales in recent years - raising the concern of a potential resurgence in transmission to humans. A campaign was therefore conducted in 2008 in order to provide dogs in South Powys with anthelmintics, estimate the coproantigen prevalence amongst these dogs, and evaluate potential risk factors for coproantigen positivity. The coproantigen prevalence identified in these dogs (n=577) was 10.6% (95% confidence interval 7.6-13.5%), with 20.6% (15.6-25.7%) of farms (n=247) containing at least one coproantigen-positive dog. Further spatial investigation identified considerable spatial heterogeneity amongst the farms, with an increased coproantigen prevalence in the regions of Brecknockshire and the Brecon Beacons National Park. Within these areas, there was strong evidence of an increase in coproantigen prevalence, compared to that found in a study conducted in 2002. Finally, a mixed effects logistic regression model identified increased roaming behaviour and decreased dog weight as being associated with increased odds of coproantigen positivity, whereas pet dogs on some farms had similar odds of infection to working dogs. Further surveillance efforts in South Powys are required and should also include the northern areas of Gwent and Glamorgan.     

Aglepristone decreases proliferation in progesterone receptor-positive canine mammary carcinomas

Background: Progesterone receptor (PR) antagonist aglepristone (RU534) has been used successfully for pregnancy termination and therapy of pyometra, vaginal tumors, and mammary hyperplasia in bitches and queens. All of these conditions share with canine mammary carcinomas the expression of PR. Objectives: To study the effect of RU534 on proliferation and apoptosis in canine mammary carcinomas in relation to PR expression. Animals: Twenty‐seven nonspayed bitches with mammary carcinomas were treated with either 2 doses of 20 mg/kg RU534 (n = 22, RU534‐treated group) or oil placebo (n = 5, control group) on days 1 and 8. Methods: Tumor samples were collected before (day 1) and after (day 15) treatment for immunohistochemistry. PR expression, proliferation index (PI), and apoptotic index (AI) were determined using antibodies against PR, Ki67, and cleaved lamin A/C antigens, respectively. The effect of treatment on these parameters was analyzed. Results: Differential expression of PR between day 1 (59.1% PR‐positive tumors) and day 15 (36.4% PR‐positive tumors) was observed in RU534‐treated tumors exclusively. After RU534 treatment, mean PI was significantly decreased in PR‐positive but unchanged in PR‐negative RU534‐treated tumors. A reduction of ≥20% in PI was found in 61.5% of RU534‐treated tumors with PR expression. Conversely, no effect on AI was observed after RU534 treatment. Conclusions and Clinical Importance: Neoadjuvant RU534 treatment had PR expression‐related inhibiting effects on proliferation of canine mammary carcinoma cells.     

Phenotypic and genetic characterization of enteropathogenic Escherichia coli (EPEC) and entero-aggregative E. coli (EAEC) from diarrhoeal and non-diarrhoeal children in Libya

A total of 50 Escherichia coli strains isolated in a Libyan hospital (20 from children with diarrhoea and 30 from healthy children) were investigated for their pathotypes and virulence traits. Altogether nine eae-positive (enteropathogenic E. coli, EPEC) and nine aggR-positive (entero-aggregative E. coli, EAEC) strains were identified. Significantly (P=0.001) more EPEC strains were identified from diarrhoeal patients (n=8) than from healthy controls (n=1), while six EAEC strains were identified from diarrhoeal and three from healthy children. Typical (eae(+), EAF(+), bfp(+)) EPEC strains (n=6) belonged to classical EPEC serogroups O55, O114, O127 and showed localized adherence on Hela cells. EAEC strains revealed genetic heterogeneity but uniformly adhered to HeLa cultures in an entero-aggregative adherence pattern. Antibiotic resistance frequently, characterized the strains. Sixty-eight percentage of the strains were resistant against at least one antibiotic and 30% harbored a class 1 integron independently of their clinical background. This is the first report from North Africa demonstrating the significance of EPEC and EAEC.     

Treatment of experimental ureteral strictures by endourological ureterotomy and implantation of stents in the porcine animal model

The objective of this study is to evaluate the dilation of the ureter using endoureterotomy and an expanding-sheath double pigtail ureteral stent in the treatment of experimentally induced ureteral strictures in the porcine animal model. This is a new treatment in the ureteral strictures resolution in Veterinary Urology, although it is not a common affection, it usually appears as a consequence of ureteritis and in the iatrogenic female genital surgery. The experimental study is design in three phases: induction of experimental stricture, diagnosis and treatment of the stricture and follow-up. We have used 10 healthy Large White female pigs. The internal ureteral diameter was measured prior to laparoscopic ligature stricture induction using retrograde ureteropyelography (RUPG). Experimental stricture was diagnosed 4 weeks after intervention, using RUPG and ultrasound, and treated by endoureterotomy and subsequent placement of a double pigtail ureteral stent, which was removed 6 weeks later. The study finished 4 weeks later with measurement of ureteral diameters using RPUG and ultrasound evaluation. Except in one case, all ureters displayed permanent dilation of the strictured area for 10 weeks after treatment (6 weeks with ureteral stent and 4 more weeks without stent). Finally, this technique proved to be effective in cases of short-length and short-living ureteral strictures, and represents a viable alternative to conventional surgery in animals.     

Leukocytic responses and intestinal mucin dynamics of broilers protected with Enterococcus faecium EF55 and challenged with Salmonella Enteritidis

The protective effect of Enterococcus faecium EF55 in chickens challenged with Salmonella enterica serovar Enteritidis phage type 4 (SE PT4) was assessed. The antibacterial effect on the bacterial microflora in the small intestine in relation to white blood cell count, phenotyping of peripheral blood and intestinal lymphocytes, functional activity of lymphocytes and phagocytes and mucin quantitation were investigated. Day-old chicks (85) were randomly divided into four groups. The probiotic group (EF) and Salmonella + probiotic group (EFSE) received E. faecium EF55 (10⁹ CFU – 3 g/group/day) for 21 days. The Salmonella group (SE) and EFSE group were infected with Salmonella Enteritidis (10⁸ CFU in 0.2 ml PBS) in a single dose per os on day four of the experiment. The control group chicks (C) were fed a commercial diet without added bacteria. Supplementation of EF55 in the diet of the chickens in the EFSE group, challenged with S. Enteritidis, caused the density of the intestinal mucin layer to increase significantly in non-specific regions (duodenum and jejunum), but decrease significantly in target regions (caeca) for S. Enteritidis. Probiotic treatment also appeared to result in a significantly higher number of lymphocytes in peripheral blood and a tendency to increase CD3, CD4, CD8, and IgM positive cells 3 days post-infection with S. Enteritidis. The results demonstrated an antibacterial effect and suggested that EF55 had a moderating effect on intestinal mucin production and leukocytic response in the early phase of S. Enteritidis infection.     

Lameness and claw lesions of the Norwegian red dairy cattle housed in free stalls in relation to environment, parity and stage of lactation

Approximately 88% of Norwegian dairy cattle are housed in tie stalls. Free stall housing for all dairy cattle will be implemented within 20 years. This means that the majority of existing stalls will be rebuilt in the near future. Fifty-seven free stall herds of the Norwegian Red breed were randomly selected and 1547 cows and 403 heifers were trimmed by 13 claw trimmers during the late winter and spring of 2002. The claw trimmers had been taught diagnosing and recording of claw lesions. Environment, management- and feeding routines were also recorded. Fifty-three herds had concrete slatted alleys while 4 had solid concrete. Thirty-five herds had concrete as a stall base, while 17 had rubber mats, 2 had wood and 3 had deep litter straw beds. The prevalence of lameness was 1.6% in hind claws. Models for lameness and claw lesions were designed to estimate the influence of different risk factors and to account for the cluster effects within herd and claw trimmer. Detected risk factors for lameness were: parity three and above and narrow cubicles; for heel horn erosions: lactation stage around 5-7 months after calving and solid concrete alleys; for haemorrhages of the white line: lactation stage around 3-5 months after calving and solid concrete alleys; for haemorrhages of the sole: parity one, lactation stage around 5-7 months after calving and short cubicles, for white line fissures: slatted concrete alleys; for asymmetrical claws: parities two and above and for corkscrewed claws: solid concrete alleys. The prevalence of lameness in heifers was low, however 29% had one or more claw lesions. Heifers that were housed in pens or free stalls had more heel-horn erosions, haemorrhages of the sole and white-line fissures than heifers in tie stalls. As new free stalls are being built, it is important to optimise the conditions for claw health.