Identification of serum stress biomarkers in pigs housed at different stocking densities

Eight Duroc×(Landrace×Large White) male pigs housed at a stocking rate of 0.50m(2)/pig were subjected to a higher stocking rate of 0.25m(2)/pig (higher density, HD) for two 4-day periods over 26 days. Using biochemical and proteomic techniques serum and plasma samples were examined to identify potential biomarkers for monitoring stress due to HD housing. HD housed pigs showed significant differences (P<0.001) in total cholesterol and low density lipoprotein-associated cholesterol, as well as in concentrations of the pig-major acute phase protein (Pig-MAP) (P=0.002). No differences were observed in serum cortisol or other acute phase proteins such as haptoglobin, C-reactive protein or apolipoprotein A-I. HD-individuals also showed an imbalance in redox homeostasis, detected as an increase in the level of oxidized proteins measured as the total plasma carbonyl protein content (P<0.001) with a compensatory increase in the activity of the antioxidant enzyme glutathione peroxidase (P=0.012). Comparison of the serum proteome yielded a new potential stress biomarker, identified as actin by mass spectrometry. Cluster analysis of the results indicated that individuals segregated into two groups, with different response patterns, suggesting that the stress response depended on individual susceptibility.     

Typical and atypical manifestations of Anaplasma phagocytophilum infection in dogs

Eighteen clinically ill dogs, naturally infected with Anaplasma phagocytophilum, were examined at a veterinary practice in Baxter, Minnesota. A clinical examination, complete blood cell count, enzyme- linked immunosorbent assay (ELISA) for A phagocytophilum, Borrelia burgdorferi, and Ehrlichia canis antibodies and Dirofilaria immitis antigen, and a polymerase chain reaction test for A phagocytophilum DNA were obtained for all dogs. Physical examination findings included fever, arthropathy, lymphadenopathy, epistaxis, acute gastritis, cervical hyperpathia, and central nervous system dysfunction. Complete blood cell count abnormalities included thrombocytopenia, morulae in neutrophils, anemia, leukopenia, eosinopenia, lymphopenia, and monocytosis. Seroreactivity to A phagocytophilum was found in 61%, B burgdorferi antibodies in 17%, and D immitis antigen in 5% of the dogs. Fever, arthropathy, neurologic dysfunction, and epistaxis are clinical syndromes that can be associated with A phagocytophilum infection. Treatment with doxycycline resulted in rapid resolution of clinical signs in all dogs.     

Genetic association between different criteria to define sexual precocious heifers with growth,carcass, reproductive and feed efficiency indicator traits in Nellore cattle using genomic information

The aim of this study was to estimate genetic parameters for different precocious calving criteria and their relationship with reproductive, growth, carcass and feed efficiency in Nellore cattle using the single‐step genomic BLUP. The reproductive traits used were probability of precocious calving (PPC) at 24 (PPC24), 26 (PPC26), 28 (PPC28) and 30 (PPC30) months of age, stayability (STAY) and scrotal circumference at 455 days of age (SC455). Growth traits such as weights at 240 (W240) and 455 (W455) days of age and adult weight (AW) were used. Rib eye area (REA), subcutaneous fat thickness (SFT), rump fat thickness (RFT) and residual feed intake (RFI) were included in the analyses. The estimation of genetic parameters was performed using a bi‐trait threshold model including genomic information in a single‐step approach. Heritability for PPC traits was moderate to high (0.29–0.56) with highest estimates for PPC24 (0.56) and PPC26 (0.50). Genetic correlation estimates between PPC and STAY weakened as a function of calving age. Correlation with SC455, growth and carcass traits were low (0.25–0.31; ?0.22 to 0.04; ?0.09 to 0.18, respectively), the same occurs with RFI (?0.09 to 0.08), this suggests independence between female sexual precocity and feed efficiency traits. The results of this study encourage the use of PPC traits in Nellore cattle because the selection for such trait would not have a negative impact on reproductive, growth, carcass and feed efficiency indicator traits. Stayability for sexual precocious heifers (PPC24 and PPC26) must be redefined to avoid incorrectly phenotype assignment.     

A comparison of the uterine proteome of mares in oestrus and dioestrus

Proteomic analysis of mare uterine flush fluid provides a minimally invasive technique for studying protein changes associated with the oestrous cycle. The aim of this study was to identify differentially abundant proteins in the uterine flush fluid of mares in oestrus and dioestrus. In this study, uterine flush fluid samples were collected from eight reproductively healthy mares in either oestrus (n = 5) or dioestrus (n = 3). Proteomic analysis was performed using liquid chromatography‐tandem mass spectrometry. Of 172 proteins identified, six proteins (immunoglobulin lambda‐like polypeptide 1, haemoglobin subunit alpha, alpha‐1B‐glycoprotein, serotransferrin, apolipoprotein A‐1, and haemoglobin subunit beta) were significantly more abundant in oestrus. These proteins may contribute to the endometrial defence system through roles in inflammation, immunity or antimicrobial activity. In other species, some of these proteins have been described as immunoglobulins, negative acute phase proteins or defence agents against micro‐organisms. During dioestrus, immunoglobulin alpha‐1 chain C region‐related, complement factor I, CD 109 antigen and uterocalin, were significantly more abundant. Research in other species suggests that these four proteins contribute to the immune response through proposed immunoregulatory characteristics, complement system involvement or roles in B cell–T cell interactions. In conclusion, ten differentially abundant proteins were identified in the uterine flush fluid of mares in oestrus and dioestrus. Targeted studies on these proteins could elucidate their role in uterine defence mechanisms during the oestrous cycle in the mare.     

Quantitative and descriptive histological aspects of jaguar (Panthera onca Linnaeus, 1758) ear skin as a step towards formation of biobanks

Skin of mammals vulnerable to extinction, such as the jaguar, is used as a source of material in conservation strategies. The composition of skin is not uniform among species, and the ability to distinguish similarities in skin morphology in animal groups is fundamental in the application of skin tissue for use in biobanks. The aim of our study was to evaluate the structure, composition and capacity for culture of ear skin from the yellow and black jaguars. Both qualitative and quantitative methods were used, focusing on skin thickness, cell quantification and distribution, collagen density, proliferative activity and viability. Histomorphometrical study of the skin showed a total thickness of 273.2 and 274.6 µm for the yellow and black jaguars, respectively. Melanocytes and fibroblasts were, respectively, 9.7 and 23.0 for the yellow jaguar and 11.3 and 26.8 for the black jaguar. A collagen density of 67.0% and 49.0% was observed for yellow and black jaguars, respectively. Both animals presented a proliferative activity varying between 1.20 and 1.30. All tissues could promote cellular detachment, reaching subconfluence in 10–15 days. This kind of information from histomorphometrical features and cell cultures can be essential for a more targeted application of ear skin cryopreservation in this species, as such information will enable understanding the action of substances on tissues during the conservation process.     

Across-breed validation study confirms and identifies new loci associated with sexual precocity in Brahman and Nellore cattle

The aim of this study was to identify candidate regions associated with sexual precocity in Bos indicus. Nellore and Brahman were set as validation and discovery populations, respectively. SNP selected in Brahman to validate in Nellore were from gene regions affecting reproductive traits (G1) and significant SNP (p ≤ 10^–3) from a meta-analysis (G2). In the validation population, early pregnancy (EP) and scrotal circumference (SC) were evaluated. To perform GWAS in validation population, we used regression and Bayes C. SNP with p ≤ 10^–3 in regression and Bayes factor ≥3 in Bayes C were deemed significant. Significant SNP (for EP or SC) or SNP in their ±250 Kb vicinity region, which were in at least one discovery set (G1 or G2), were considered validated. SNP identified in both G1 and G2 were considered candidate. For EP, 145 SNP were validated in G1 and 41 in G2, and for SC, these numbers were 14 and 2. For EP, 21 candidate SNP were detected (G1 and G2). For SC, no candidate SNP were identified. Validated SNP and their vicinity region were located close to quantitative trait loci or genes related to reproductive traits and were enriched in gene ontology terms related to reproductive success. These are therefore strong candidate regions for sexual precocity in Nellore and Brahman.     

Different isolates from Leishmania braziliensis complex induce distinct histopathological features in a murine model of infection

The aim of this study was to evaluate the histopathological features in tissues of mice infected by human isolates (I, II, and III) or the reference M2903 strain of Leishmania braziliensis complex. BALB/c and C57Bl/6 mice were infected in the hind footpad with 10⁶ stationary-phase promastigotes of L. braziliensis complex. The evolution of lesions was observed for 10 weeks and the animals were then euthanized and liver, spleen and popliteal lymph nodes were collected. Tissues were stained with hematoxylin and eosin and analyzed by immunohistochemistry assay. Increased thickness of infected footpads was observed in all animals, lesions were nodular and non-ulcerated. Mice infected with isolate I presented inflammatory infiltrates consisting predominantly of mononuclear cells in all tissues examined, and also a great number of megakaryocytes, compared with other isolates. Infection with isolate II led to an infected footpad enlargement not seen in other isolates. In addition, mononuclear infiltrates in the liver and hemosiderin in spleen were noted. Conversely, mice infected with either isolate III or M2903 strain only showed an increased number of megakaryocytes in spleen. All tissues examined had detectable amastigote forms of Leishmania by immunohistochemistry in all groups. Taking together, our results showed an unforeseen behavior of different isolates of L. braziliensis complex that led to diverse pathological findings.     

Ixodid ticks parasitizing Iberian red deer (Cervus elaphus hispanicus) and European wild boar (Sus scrofa) from Spain: geographical and temporal distribution

Commercial hunting of Spanish wild ungulates has made them an important economic resource. Wild ungulates may have an important role in the maintenance of ixodid tick populations, and also as reservoirs of pathogens. We studied the ixodid ticks that parasitize Iberian red deer and European wild boar from Spain. Ixodid ticks (n=6,336) were collected from 431 Iberian red deer and 142 wild boar in different regions of Spain. We found 10 different ixodid tick species parasitizing Iberian red deer, mainly Hyalomma marginatum marginatum (63.7%), Rhipicephalus (Boophilus) annulatus (7.9%) and R. bursa (7.5%). R. (Boophilus) annulatus was only collected in the province of Cádiz (southern Spain). We found 8 ixodid tick species on the wild boar, mainly Hy. m. marginatum (68.7%), R. bursa (14.6%) and Dermacentor marginatus (9.3%). We found one adult Hy. marginatum rufipes and one adult Hy. anatolicum excavatum parasitizing wild boar from south-central Spain. Mean prevalence of ixodid ticks was 41.3+/-0.08% (n=475) and 31+/-0.09% (n=284) and intensity of parasitization was 13.9+/-0.2 (n=283) and 13.6+/-0.3 (n=130) ticks/animal for Iberian red deer and wild boar, respectively. Only 5 of the 13 ixodid tick species found were shared by Iberian red deer and wild boar. This finding could indicate a host preference when Iberian red deer and wild boar share common habitats. In both Iberian red deer and wild boar from south-central Spain the monthly relative frequencies of Hy. m. marginatum and R. bursa presented an inverse pattern. The highest Hy. m. marginatum relative frequencies coincided with the lowest R. bursa relative frequencies along the year. R. bursa and I. ricinus were present in areas from northern to southern Spain while Hyalomma sp. and D. marginatus were exclusively collected in the two southern thirds of Spain. Haemaphysalis sp. and D. reticulatus were collected in northern Spain. Hy. m. marginatum and R. bursa were present during the whole year in red deer and wild boar from south-central Spain, showing more than one life cycle per year. These results are important for understanding the role of wild ungulates in the maintenance of tick infestations and to improve tick control programmes.     

Intracranial fusariosis: a novel cause of fungal meningoencephalitis in a dog

The Fusarium species are a group of saprophytic fungal organisms that are occasionally the cause of opportunistic infections in humans and animals. Central nervous system disease associated with a Fusarium species is most commonly described in horse, resulting in equine leukoencephalomalacia. This report describes a 2-year-old, spayed, female German Shepherd Dog with meningoencephalitis secondary to infection with Fusarium spp. Meningoencephalitis in dogs secondary to a species of Fusarium has not been previously reported. The diagnosis was made based on the histopathologic examination of brain tissues postmortem and special immunohistochemical stains specific for Fusarium solani. The clinical signs in this dog were indicative of multifocal brain disease and included seizures and a paradoxical vestibular syndrome. The clinical findings, diagnostic and histopathologic test results, and the comparative characterizations of other disseminated fungal diseases, especially aspergillosis, are described.     

Hepatozoon canis infection associated with dog ticks of rural areas of Rio de Janeiro State, Brazil

Hepatozoon canis is a tick-borne protozoan that infects dogs and has been reported throughout the world. Manifestation of H. canis infection varies from being sub-clinical in apparently healthy dogs to severe illness. The main vector of the infection is the dog tick, Rhipicephalus sanguineus although other species may also transmit this agent. H. canis has been reported previously in Brazil, but mostly as an occasional finding during laboratory exams and always associated with other diseases. The prevalence of H. canis in dogs of rural areas of Brazil has been little studied. For this study, 250 dogs from seven counties of Rio de Janeiro state were examined. All the dogs were from rural areas, near forest. Of the dogs examined, 26 dogs were from Seropédica, 82 from Itaguaí, 41 from Paracambi, 26 from Mangaratiba, 32 from Barra do Piraí, 32 from Piraí and 11 from Miguel Pereira. Blood smears from the peripheral blood of the ear were taken and ticks found on the dogs were collected for identification in the laboratory. Using blood smear evaluation, H. canis was identified in 39.2% of the animals examined. Other hemoparasites identified were Babesia canis (5.2%) and Ehrlichia canis (4.8%). Four tick species were found parasitizing the dogs: Amblyomma cajennense (23.6%), R. sanguineus (12.4%), Amblyomma aureolatum (2.8%) and Amblyomma ovale (2.0%). There was a positive correlation between the presence of A. cajennense and H. canis infection.