Endostatin in 3D Fibrin Hydrogel Scaffolds Promotes Chondrogenic Differentiation in Swine Neonatal Meniscal Cells.

The success of cell-based approaches for the treatment of cartilage or fibro-cartilaginous tissue defects requires an optimal cell source with chondrogenic differentiation ability that maintains its differentiated properties and stability following implantation. For this purpose, the aim of this study was to evaluate the use of endostatin (COL18A1), an anti-angiogenic factor, which is physiologically involved in cell differentiation during meniscus development. Swine neonatal meniscal cells not yet subjected to mechanical stimuli were extracted, cultured in fibrin hydrogel scaffolds, and treated at two different time points (T1 = 9 days and T2 = 21 days) with different concentrations of COL18A1 (10 ng/mL; 100 ng/mL; 200 ng/mL). At the end of the treatments, the scaffolds were examined through biochemical, molecular, and histochemical analyses. The results showed that the higher concentration of COL18A1 promotes a fibro-chondrogenic phenotype and improves cellularity index (DNA content, p < 0.001) and cell efficiency (GAGs/DNA ratio, p < 0.01) after 21 days. These data are supported by the molecular analysis of collagen type I (COL1A1, a marker of fibrous-like tissue, p < 0.001), collagen type II (COL2A1, a marker of cartilaginous-like tissue, p < 0.001) and SRY-Box Transcription Factor 9 (SOX9, an early marker of chondrogenicity, p < 0.001), as well as by histological analysis (Safranin-O staining), laying the foundations for future studies evaluating the involvement of 3D endostatin hydrogel scaffolds in the differentiation of avascular tissues.

Does Immunocastration Affect Behaviour and Body Lesions in Heavy Pigs?

Immunocastration has been pointed out as an alternative to surgical castration; though, most of the scientific studies were performed in light pig production. This study aimed to evaluate the effect of immunocastration on animal welfare in heavy pig production through the evaluation of behaviour and body lesions. A total of 188 commercial-hybrid pigs were randomly allocated into two treatment groups: surgical castration (SC) and immunocastration with Improvac® (IC). Data on behaviour, body lesions, and salivary testosterone levels were collected the day before each vaccination at 15, 22, 32, and 36 weeks of age. IC and SC pigs were slaughtered at 40 and 41 weeks of age, respectively; productive and carcass traits data were also collected. Considering productive performance, our results confirmed that IC pigs grew faster and presented a higher weight at slaughter. A critical period for pig welfare was observed before 32 weeks: testosterone concentration and body lesion score were significantly higher in IC pigs compared to SC pigs; active behaviours were significantly more frequent in IC at 15 weeks. Immunocastration may represent a suitable alternative to surgical castration with profitable productive performances, whereas the impairment of welfare during the period before the effective vaccination should be further investigated as a potential critical aspect in heavy pig production.

A survey of Italian cat owners' attitudes towards cat vaccination through a web-based questionnaire.

BACKGROUND: Vaccination plays an important role in feline healthcare as it is the most effective measure for prevention against feline infectious diseases. Therefore, it is important to know owners' opinion towards cats' vaccination and current veterinary practices in order to advice owners on the use of the correct vaccination protocol. This study aimed to investigate the proportion of cats regularly vaccinated and identify the main factors motivating cat owners' decisions related to vaccination in Italy. A questionnaire was disseminated online (mainly via social networks) to collect data regarding Italian cat owners' demographics, information about cats, factors regarding cats' vaccination, and veterinary-owner relationship. RESULTS: A total of 1264 owners participated in the survey and 1247 questionnaires were completed and statistically analyzed. The majority (91%; n = 1131) of cats were vaccinated and 80% (n = 998) had been vaccinated within the last 3 years. Age of 2-4 years old cats and the acquisition from a breeder or cat shop were significantly associated with vaccination within the last 3 years. Cats vaccinated but not within the last 3 years were significantly associated with cat's indoor lifestyle, cats' age ≥ 5 years old and low annual household income. Importance of vaccination cost, low annual household income and owners' job not related to healthcare was statistically associated with the lack of cat's vaccination. In addition, 86% of the owners took their cat regularly to veterinary clinics. Veterinarians play a significant role in owners' decision, and they are considered the most useful source of information about vaccination by 97% of owners. CONCLUSIONS: The high number of recently vaccinated cats suggests owner's attention towards feline vaccination and cat's health. The importance of veterinarian's advice along with the knowledge of factors associated to the unvaccinated status of cats may help veterinarians to grow owner's confidence and increase prevention of feline infectious diseases. However further investigations based on a more comprehensive sample of the general population are needed to confirm the results of this survey.

Translational Approach to Induce and Evaluate Verocytotoxic E. coli O138 Based Disease in Piglets.

Pig livestock was influenced by several global concerns that imposed a re-thinking of the farming system, which included the reduction in chemical dependency and the development of antimicrobial alternatives. Post-weaning diarrhea and enterotoxaemia caused by Escherichia coli, are serious threats that are responsible for the economic losses related to mortality, morbidity and stunted growth in weaning piglets. The aim of the study was to set up experimental conditions to simulate the simultaneous outbreak of post-weaning diarrhea and enterotoxaemia in weaned piglets, through verocytotoxic O138 Escherichia coli challenge, with a multidisciplinary approach. Eighteen piglets susceptible to F18 VTEC infection were selected by polymerase chain reaction for polymorphism on the fucosyltransferase 1 gene and randomly divided in two experimental groups, non-infected controls (C; n = 6) and infected ones (I; n = 12) and housed into individual pens at the same environmental conditions for 29 days. At day 20, I pigs were orally inoculated with Escherichia coli O138 and fed a high protein ration for 3 days. Zootechnical, clinical, microbiological, histological and immunological parameters were evaluated along the follow up (3 and 9 days). Experimental infection, confirmed by bacteria faecal shedding of the I group, significantly affected the clinical status. The I group showed significantly higher total scores, corresponding to medians of the sum of daily scores from days 1 to 3 (Σ3) and 1 to 9 (Σ9) post infection, epiphora, vitality, hair irregularity, oedema and depression. Histological examination showed evident inflammatory infiltrate of lymphocytes, and follicular hyperplasia in I pigs; in the same group, the immunohistochemical and immunological assays revealed an increase in IgG in the intestinal crypts and CD3-positive T cells in intestinal epithelium. The experimental Escherichia coli infection in controlled conditions is crucial for both the evaluation of innovative compounds and the elucidation of the mechanisms associated with the persistence of antibacterial resistant strains. In conclusion, the adopted infection model, carried out on receptor-mediated susceptible piglets, allowed us to identify a discriminative panel of clinical symptoms related to Escherichia coli O138 infection, and could be used to assess the protective effect of antibiotic alternatives.

Innate immune responses to metabolic stress can be detected in rumen fluids.

Many production diseases of dairy cows are related to digestive troubles. The rumen subacute acidosis is the most relevant one, albeit not easily recognized. Recent studies suggest that forestomachs can perform regulatory actions at both regional and systemic levels, since forestomach walls express immune receptors and cytokines, and the rumen liquor is infiltrated by leukocytes. Therefore, the rumen fluid could be conveniently collected for investigating metabolic production diseases. Thus, we investigated the origin of the leukocytes of the rumen fluid and demonstrated that they partly derive from saliva. Next, we carried out a field survey of innate immunity in rumen fluids of 128 cows from 12 dairy farms, along with clinical inspections, assessment of milk yield, rumen pH, volatile fatty acids (VFA) and major inflammo-metabolic parameters. Significant statistical correlations were found between immune markers in rumen fluids and biochemical parameters. A significant negative correlation was found in rumen between CD45 gene expression (leukocyte infiltration) and pH level. B cells were the most frequent mononuclear leukocyte population in the rumen liquor and their infiltration was negatively affected by low ruminal pH and high concentrations of VFA. Moreover, total Ig and IgM in rumen fluids were negatively correlated with ruminal pH and positively correlated with uremia. Our data suggest that forestomach immune responses could be directed to "dangers" arising within the forestomach environment. The immune markers could integrate consolidated diagnostic parameters (e.g. rumen pH) and contribute to robust, early diagnosis of tricky digestive troubles of cattle.