Ejaculation does not contribute to the stress response to electroejaculation in sheep

Electroejaculation procedures (EEPs) provoke stress; nevertheless, ejaculation produces physiological changes similar as those usually used to measure stress responses. The application of EEP to animals that cannot ejaculate—as ewes—may be useful to discriminate the responses induced by ejaculation from those provoked by EEP. The aim was to determine the stress response to EEP in rams and ewes. The EEPs were applied to 10 rams and 10 ewes during the non‐breeding season, and the number of vocalizations, the heart rate, rectal temperature, serum cortisol concentration, biochemical and haematological parameters were measured. Overall, EEP provoked increases in cortisol concentration, glycaemia, rectal temperature and concentration of creatine kinase (all them: p < .0001) as well as relative concentration of granulocytes (p = .003) and absolute granulocyte concentration (p = .0002) in both, rams and ewes. Heart rate, relative concentration of lymphocytes (p = .001), haematocrit (p = .02) and haemoglobin (p = .045) decreased in animals from both genders after EEP. Besides, cortisol (p < .0001), rectal temperature (p = .002) and glycaemia (p = .001) were greater in ewes than rams, and creatine kinase also tended to be greater in ewes than rams (p = .054). On the other hand, the number of animals that vocalized (p = .006), white blood cells (p = .02) and absolute lymphocytes (p = .02) were greater in rams than ewes. The general trends show a similar pattern of stress responses in animals from both genders. Therefore, we concluded that ejaculation does not contribute to the stress response provoked by the EEP. This procedure also provokes muscular damage and probably pain.     

Twelfth rib resection as an approach for portal vein cannulation in sheep.

A surgical technique involving resection of the twelfth rib was used to insert silastic cannulas into the portal veins of three sheep to study amino acid metabolism. Good exposure to the vein was achieved by this method although it required positive ventilation due to the penetration of the thoracic cavity. All cannulas were buried subcutaneously and exteriorized near the dorsal midline. This facilitated continuous infusion into the portal cannula without disturbing cannula placement.     

Pathogenesis of acute toxoplasmosis in specific-pathogen-free cats

Systemic toxoplasmosis was produced in specific-pathogen-free cats by intravenous inoculation with Toxoplasma gondii tachyzoites. Infectious organisms were recovered from all tissues studied, but the number of organisms recovered from liver, lungs and spleen was 10-fold to 10,000-fold higher than from heart and brain. The occurrence and severity of Toxoplasma-induced lesions correlated with the number of infectious organisms recovered from the various tissues. In nonlymphoid tissues, the Toxoplasma-associated lesions consisted of multifocal necrosis, usually with demonstrable organisms. Lesions in the spleen and mesenteric lymph nodes consisted of reticuloendothelial and lymphoid hyperplasia, with few demonstrable organisms. Pneumonitis was severe and sometimes fatal in the early stages of systemic toxoplasmosis. Light- and electron-microscopic studies showed that the earliest lung lesions were randomly distributed infiltrates of neutrophils, eosinophils, and mononuclear cells into alveolar walls. Later lesions were diffuse alveolar necrosis, pneumocytic hyperplasia, and extensive fibrinocellular exudates in alveoli. Tachyzoites were present in cytoplasmic vacuoles of fibroblasts, macrophages, type I and II pneumocytes, bronchiolar epithelial cells, bronchiolar smooth muscle cells, endothelial cells, neutrophils, and eosinophils, and circulating monocytes. Replication of organisms was found in all parasitized cell types except neutrophils and eosinophils.     

IgG from acutely infected cats blocks mucosal feline immunodeficiency virus infection

We have previously shown an absence of detectable systemic or local infection in cats exposed to an infectious (100 TCID(50)) feline immunodeficiency virus (FIV) plasma inoculum via either the rectal or vaginal mucosa. In contrast, this same plasma inoculum was infectious via parenteral inoculation. Moreover an equivalent dose of cell-free tissue culture-origin virus inoculum infected 100% of cats by either the rectal or vaginal exposure route. To evaluate this phenomena, we used a tissue culture system to identify a heat-stable factor in the plasma of cats acutely (3 weeks) infected with FIV that blocked infection of naive peripheral blood mononuclear cells (PBMC) by either cell-free or cell-associated FIV in vitro. A single application of as little as a 1:200 dilution of either heparinized or Alsevier's anticoagulated plasma effectively inhibited production of FIV p26 in culture over a 21-day co-culture period. Depletion of antibody using a protein A column abrogated the inhibitory effect of FIV plasma against in vitro FIV infection. Co-inoculation of heat-inactivated plasma with 400 TCID(50) FIV-B-2542 cell-free supernatant virus onto the vaginal mucosa of two cats resulted in complete inhibition of infection in one cat and increased time to infection in the second. Thus, antibody found in the plasma of cats acutely infected with FIV blocks cell-associated and cell-free infection, inhibits virus production in previously infected cells, and reduces mucosal transmission efficiency in vivo. Extrapolation may help explain the relatively inefficient mucosal transmission of human immunodeficiency virus-1 (HIV) and other lentiviruses.     

The immune response and maternal antibody interference to a heterologous H1N1 swine influenza virus infection following vaccination

This study investigated the efficacy of a bivalent swine influenza virus (SIV) vaccine in piglets challenged with a heterologous H1N1 SIV isolate. The ability of maternally derived antibodies (MDA) to provide protection against a heterologous challenge and the impact MDA have on vaccine efficacy were also evaluated. Forty-eight MDA(+) pigs and 48 MDA(-) pigs were assigned to 8 different groups. Vaccinated pigs received two doses of a bivalent SIV vaccine at 3 and 5 weeks of age. The infected pigs were challenged at 7 weeks of age with an H1N1 SIV strain heterologous to the H1N1 vaccine strain. Clinical signs, rectal temperature, macroscopic and microscopic lesions, virus excretion, serum and local antibody responses, and influenza-specific T-cell responses were measured. The bivalent SIV vaccine induced a high serum hemagglutination-inhibition (HI) antibody titer against the vaccine virus, but antibodies cross-reacted at a lower level to the challenge virus. This study determined that low serum HI antibodies to a challenge virus induced by vaccination with a heterologous virus provided protection demonstrated by clinical protection and reduced pneumonia and viral excretion. The vaccine was able to prime the local SIV-specific antibody response in the lower respiratory tract as well as inducing a systemic SIV-specific memory T-cell response. MDA alone were capable of suppressing fever subsequent to infection, but other parameters showed reduced protection against infection compared to vaccination. The presence of MDA at vaccination negatively impacted vaccine efficacy as fever and clinical signs were prolonged, and unexpectedly, SIV-induced pneumonia was increased compared to pigs vaccinated in the absence of MDA. MDA also suppressed the serum antibody response and the induction of SIV-specific memory T-cells following vaccination. The results of this study question the effectiveness of the current practice of generating increased MDA levels through sow vaccination in protecting piglets against disease.     

In vivo CXCR4 expression, lymphoid cell phenotype, and feline immunodeficiency virus infection

Primary isolates of feline immunodeficiency virus (FIV) appear to require binding to CD134 in conjunction with CXCR4(X4) to infect IL-2-dependent T-cell-derived cells in culture. However, much less is known about the role of X4 for the infection of cells in vivo. To investigate the correlation between X4 expression and FIV infection in cats acutely infected with FIV-C-Pgmr we used high-speed fluorescence-activated cell sorting and realtime PCR to co-analyze cell phenotypes from lymph node, thymus, bone marrow and blood for FIV infection and X4 expression. X4 expression was greatest in lymph node, both in frequency and in mean fluorescence intensity. The thymus demonstrated a higher proviral burden in X4+ thymic T cells (14% in X4+ thymic T cells and 7% in X4− cells) whereas, proviral loads were similar between X4+ and X4− cell populations in all other tissues examined. Assuming a minimum of one proviral copy per cell, a maximum of 50% of FIV-positive cells were X4+. The highest fraction of FIV-infected X4− cells was present in bone marrow. Regardless of X4 status, proviral loads were higher in lymph node and blood T cells than in B cells. These studies provide both a positive association between X4 expression and FIV infection and introduce the probability that X4-independent infection occurs in other target cells in vivo.