Effects of maternal subnutrition during early pregnancy on cow hematological profiles and offspring physiology and vitality in two beef breeds

This experiment evaluated the effects of subnutrition during early gestation on hematology in cows (Bos Taurus) and on hematological, metabolic, endocrine, and vitality parameters in their calves. Parda de Montaña and Pirenaica dams were inseminated and assigned to either a control (CONTROL, 100% requirements) or a nutrient‐restricted group (SUBNUT, 65%) during the first third of gestation. Dam blood samples were collected on days 20 and 253 of gestation, and calf samples were obtained during the first days of life. Pirenaica dams presented higher red series parameters than Parda de Montaña dams, both in the first and the last months of gestation. During early pregnancy, granulocyte numbers and mean corpuscular hemoglobin were lower in Pirenaica‐SUBNUT than in Pirenaica‐CONTROL cows. Calves from the SUBNUT cows did not show a physiological reduction in red series values in early life, suggesting later maturation of the hematopoietic system. Poor maternal nutrition affected calf endocrine parameters. Newborns from dystocic parturitions showed lower NEFA concentrations and weaker vitality responses. In conclusion, maternal nutrition had short‐term effects on cow hematology, Pirenaica cows showing a higher susceptibility to undernutrition; and a long‐term effect on their offspring endocrinology, SUBNUT newborns showing lower levels of IGF‐1 and higher levels of cortisol.     

Home range variability,spatial aggregation,and excursions of Akodon azarae and Oligoryzomys flavescens in Pampean agroecosystems

Rodents are reservoirs of various types of hantavirus, some of which are agents of hantavirus pulmonary syndrome in humans. Each hantavirus is associated with a single rodent host species but successive spill‐over events may eventually lead to host‐switching and new species’ becoming host of a given pathogen. This study aims to gain an understanding of the spatial ecology of two hantavirus‐host species, Akodon azarae, and Oligoryzomys flavescens, by identifying factors modulating their home range sizes and stability, and by evaluating intra‐ and interspecific spatial aggregation for these species and a third one—Oxymycterus rufus—living in sympatry. For this, eleven capture‐mark‐recapture surveys were carried out, spanning 22 months. We found that A. azarae males have larger and more mobile home ranges than females, independently of the season. Consequently, males could likely have a more relevant role in the transmission of hantavirus because of their greater exposure both to a higher number of contacts between individuals and viral contamination of the environment. Contrasting, O. flavescens individuals showed negligible displacements of their home range through time, which could limit the range of hantavirus spread in host populations. Since O. flavescens is host to Lechiguanas hantavirus (pathogenic to humans) this result encompasses epidemiological relevance, for it may imply the existence of local foci of infection. Additionally, individuals of both species performed excursions outside their home ranges. These events could enable hantavirus spread over distances beyond the normal range of movements and lead to new hantavirus outbreaks in formerly non‐infected rodent populations, favoring the persistence of the virus in nature.     

Autophagic response in the Rabbit Hemorrhagic Disease,an animal model of virally-induced fulminant hepatic failure

The Rabbit Hemorrhagic Disease Virus (RHDV) induces a severe disease that fulfils many requirements of an animal model of fulminant hepatic failure. However, a better knowledge of molecular mechanisms contributing to liver damage is required, and it is unknown whether the RHDV induces liver autophagy and how it relates to apoptosis. In this study, we attempted to explore which signalling pathways were involved in the autophagic response induced by the RHDV and to characterize their role in the context of RHDV pathogenesis. Rabbits were infected with 2 × 10⁴ hemmaglutination units of a RHDV isolate. The autophagic response was measured as presence of autophagic vesicles, LC3 staining, conversion of LC3-I to autophagosome-associated LC3-II and changes in expression of beclin-1, UVRAG, Atg5, Atg12, Atg16L1 and p62/SQSTM1. RHDV-triggered autophagy reached a maximum at 24 hours post-infection (hpi) and declined at 30 and 36 hpi. Phosphorylation of mTOR also augmented in early periods of infection and there was an increase in the expression of the endoplasmic reticulum chaperones BiP/GRP78, CHOP and GRP94. Apoptosis, measured as caspase-3 activity and expression of PARP-1, increased significantly at 30 and 36 hpi in parallel to the maximal expression of the RHDV capsid protein VP60. These data indicate that RHDV infection initiates a rapid autophagic response, perhaps in an attempt to protect liver, which associates to ER stress development and is independent from downregulation of the major autophagy suppressor mTOR. As the infection continues and the autophagic response declines, cells begin to exhibit apoptosis.     

Testing the effectiveness of conservation management within biosphere reserves: the case of the Mexican deer mouse (Peromyscus mexicanus) as a bioindicator

Biosphere reserves are protected areas whose purpose is to combine conservation and sustainable development. However, their effectiveness has not been tested sufficiently, especially from an ecological and genetic approach. In this sense, the Peromyscus genus represents an excellent bioindicator to address these questions, due to its short life and high evolutionary rate and fecundity. For conservation managers, genetic structure can increase the rate of loss of genetic diversity because alleles exclusive of a subpopulation are more likely to disappear as a consequence of genetic drift in comparison with a panmictic population. Here we analyzed the abundance, movement distances, morphology/morphometry and genetic structure of 3 populations of Mexican deer mouse (Peromyscus mexicanus) located in different protected zones of La Tigra National Park (Honduras). Our results are consistent among the 3 approaches and showed the highest values of abundance, morphometry and genetic diversity in the population located at the core zone, whereas non‐statistically significant differences were found between buffer and transition zone populations, suggesting suitable effectiveness of conservation management in the core zone but a lack of ecological buffering function of the other zones. In addition, the low movement distances and high genetic structure among the studied populations provide evidence of poor conservation management in the buffer and transition zone. Thus, we discuss the utility of the novel methodology used in this work, combining morphometry, abundance and genetics, in testing the effectiveness of conservation strategies in biosphere reserves, and the value of the Peromyscus genus as a bioindicator.     

Immunohistochemical detection of extrinsic and intrinsic mediators of apoptosis in porcine paraffin-embedded tissues

Apoptosis is a strictly regulated mechanism of cell death that involves a complex network of biochemical pathways. Whether a cell undergoes apoptosis or not depends on a delicate balance of anti- and pro-apoptotic stimuli. This phenomenon can be induced by two different pathways: intrinsic and extrinsic pathways. The main aim of this study was to determine the ideal fixative and antigen retrieval method in porcine paraffin embedded tissues for the immunohistochemical detection of apoptosis mediators, from both extrinsic and intrinsic pathways. Tonsil, retropharyngeal lymph node and lung tissue samples were fixed in 10% neutral buffered formalin, Bouin solution and zinc salts fixative (ZSF) and different unmasking methods were carried out. Both 10% neutral buffered formalin and ZSF resulted as the fixatives of election to study apoptosis phenomena. Tween 20 (0.01% in PBS), citrate buffer (microwave, pH 6.0) and/or protease type XIV were the antigen retrieval methods which displayed better labelling. Our results allow to deep in the knowledge of apoptosis and its role in the pathogenesis of porcine diseases.     

Radiation up-regulates the expression of VEGF in a canine oral melanoma cell line

To evaluate radiosensitivity and the effects of radiation on the expression of vascular endothelial growth factor (VEGF) and VEGF receptors in the canine oral melanoma cell line, TLM 1, cells were irradiated with doses of 0, 2, 4, 6, 8 and 10 Gray (Gy). Survival rates were then determined by a MTT assay, while vascular endothelial growth factor receptor (VEGFR)-1 and -2 expression was measured by flow cytometry and apoptotic cell death rates were investigated using an Annexin assay. Additionally, a commercially available canine VEGF ELISA kit was used to measure VEGF. Radiosensitivity was detected in TLM 1 cells, and mitotic and apoptotic cell death was found to occur in a radiation dose dependent manner. VEGF was secreted constitutively and significant up-regulation was observed in the 8 and 10 Gy irradiated cells. In addition, a minor portion of TLM 1 cells expressed vascular endothelial growth factor receptor (VEGFR)-1 intracellularly. VEGFR-2 was detected in the cytoplasm and was down-regulated following radiation with increasing dosages. In TLM 1 cells, apoptosis plays an important role in radiation induced cell death. It has also been suggested that the significantly higher VEGF production in the 8 and 10 Gy group could lead to tumour resistance.     

Seroprevalence for norovirus genogroups GII and GIV in captive non‐human primates

Noroviruses (NoVs) are a major cause of epidemic gastroenteritis in children and adults. Several pieces of evidence suggest that viruses genetically and antigenically closely related to human NoVs might infect animals, raising public health concerns about potential cross‐species transmission. The natural susceptibility of non‐human primates (NPHs) to human NoV infections has already been reported, but a limited amount of data is currently available. In order to start filling this gap, we screened a total of 86 serum samples of seven different species of NPHs housed at the Zoological Garden (Bioparco) of Rome (Italy), collected between 2001 and 2017, using an enzyme‐linked immunosorbent assay (ELISA) based on virus‐like particles (VLPs) of human GII.4 and GIV.1 NoVs. Antibodies specific for both genotypes were detected with an overall prevalence of 32.6%. In detail, IgG antibodies against GII.4 NoVs were found in 18 Japanese macaques (29.0%, 18/62), a mandrill (10.0%, 1/10), a white‐crowned mangabey (16.6%, 1/6) and in an orangutan (33.3%, 1/3). Twelve macaques (19.3%, 12/62), five mandrills (50.0%, 5/10), two chimpanzees (100%, 2/2) and a white‐crowned mangabey (16.6%, 1/6) showed antibodies for GIV.1 NoVs. The findings of this study confirm the natural susceptibility of captive NHPs to GII NoV infections. In addition, IgG antibodies against GIV.1 were detected, suggesting that NHPs are exposed to GIV NoVs or to antigenically related NoV strains.     

Porcine reproductive and respiratory syndrome virus (PRRSV)-specific mAbs: supporting diagnostics and providing new insights into the antigenic properties of the virus

The porcine reproductive and respiratory syndrome virus (PRRSV) is one of the most important viral pathogens in the swine industry. Despite great efforts of pig holders, veterinarians, researchers and vaccine developers, the virus still causes major production losses. It is clear that efficient and correct monitoring and rational development of vaccines are crucial in the combat against this pathogen. PRRSV-specific monoclonal antibodies (mAbs) are essential tools for both diagnostic and research purposes. This study describes the production of PRRSV GP3-, GP5- and N-specific hybridomas and an extensive characterization of the mAbs. The N-specific mAbs generated in this study appear to be useful tools for diagnostics, as they were found to react with genetically very different PRRSV isolates and may serve to discriminate between European and American type PRRSV isolates. These mAbs also allowed detection of the PRRSV N protein in both formalin-fixed, paraffin-embedded tissue sections and frozen tissue sections of PRRSV-infected lungs, further illustrating their diagnostic value. Different neutralization assays pointed out that none of the GP3- and GP5-specific mAbs tested shows virus-neutralizing capacity. This is noteworthy, as these mAbs recognize epitopes in the predicted ectodomains of their target protein and since the GP5-specific antibodies specifically react with the antigenic region that corresponds to the "major neutralizing epitope" suggested for American type PRRSV. The current findings argue against an important role of the identified antigenic regions in direct antibody-mediated neutralization of European type PRRSV in vivo. However, it is also clear that findings concerning a specific PRRSV epitope cannot always be generalized, as the antigenic determinants and their biological properties may differ radically between different virus isolates.     

Transmission dynamics of Fasciola hepatica in the Plateau Region of Mexico. Effect of weather and treatment of mammals under current farm management

The aim of the present work was to study the dynamics of Fasciola hepatica natural infection in ovines, caprines, bovines and two mollusc species, Lymnaea (Fossaria) humilis and Lymnaea (F.) bulimoides, from 2004 to mid 2007 under normal farm management conditions, and the relation to climate changes. The study was performed in a research centre in the plateau of Mexico. Temperature and rainfall were registered every month, as well as the number and intensity of infection in livestock and molluscs, as determined by coprology and direct observation/cercariae release, respectively. The first two years mammals were treated with clorsulon/ivermectin because the animals were harbouring concomitant intestinal nematode infections and this was the available drug combination. During the second period treatment was with triclabendazole. The temperature ranged from around cero to 30 °C, except during September 2005 to January 2006, when a cold climate prevailed. The rainfall augmented every year in July-August, and slightly in April, 2006. Lymneid snails appeared during or immediately after the rainfall peaks of 2004 and 2006, while few L. humilis and no L. bulimoides were present during the same period of 2005, probably because it was cold. A total of 15564 cercariae were released from molluscs during the wet time of 2004, 76 during 2005 and 368 in 2006. Several peaks of infection in mammals were observed, most occurring up to 4 months after the snails had disappeared. As expected, the weather had strong impact on snails and then on livestock infection. Also, treatment given to livestock was related to reduced cercarieae release five months later. Therefore, the combination of treatment and inspection of snails in the biotopes where the livestock graze may facilitate control of fasciolosis under current farm management.     

Signaling pathways involved in liver injury and regeneration in rabbit hemorrhagic disease, an animal model of virally-induced fulminant hepatic failure

Management of fulminant hepatic failure (FHF) continues to be one challenging problem, and experimental animal models resembling its clinical conditions are still needed. Rabbit hemorrhagic disease (RHD) fullfils many requirements of an animal model of FHF. This work investigated changes in MAPK, NF-κB, AP-1 and STAT pathways during RHD-induced liver injury. Rabbits were infected with 2 × 10⁴ hemagglutination units of an RHD virus isolate. Apoptosis was documented by the presence of caspase-3 activity and substantial PARP proteolysis at 36 and 48 h postinfection (pi). Infection induced a marked and maintained expression of TNF-α from 12 h pi, while there was only a transitory increase in IL-6 expression. Expression of phosphorylated (p)-JNK, p-p38 and p-ERK1/2 was significantly elevated at 12 h pi. At 48 h pi p-JNK expression was maintained at a maximum level, while that of p-p38 returned to normality and there was no p-ERK1/2 expression. Activation of NF-κB and AP-1 and increased expression of VCAM-1 and COX-2 were observed. No significant changes were detected in activation of STAT1 and STAT3, while SOCS3 expression increased significantly. The current findings suggest that activation of JNK is an essential component in liver injury mediated by the RHD virus and that lack of activation of STAT3, probably mediated by SOCS3 over-expression, would contribute to the inhibition of the regenerative response. Data show the presence of molecular mechanisms contributing to liver damage and the lack of regeneration and they support the usefulness of this model to investigate novel therapeutical modalities in FHF.