Molecular detection of Tritrichomonas foetus in cats from Abruzzo region (central Italy)

INTRODUCTION. Tritrichomonas foetus (Trichomonadida, Tritrichomonadidae), a protozoan parasite causing chronic diarrhea in domestic cats and transmitted through the fecal-oral route (Gookin et al., 2004J Clin Microbiol. 42: 2707-10), has been reported worldwide. The clinical manifestations caused by T. foetus are variable, ranging from subclinical to severe. The infection is more frequent in catteries and shelters, and in purebred and young cats.Only few studies have investigated the occurrence of T. foetus in catsfrom limited regions of Italy (Mancianti et al., 2015J Feline Med Surg. 17: 163-67; Veronesi et al., 2016 Vet Parasitol Reg Stud Reports. 6: 14-19). This study has investigated its prevalence in cats froma region of Central Italy. MATERIALS AND METHODS. In 2019, individual faecal samples were collected from105 cats (37 privately owned, 68 colony-cats) referred at the University Veterinary Teaching Hospital of Teramo, Abruzzo region. The faecal samples were tested using aspecific PCR protocol (Gookin et al., 2002 J Clin Microbiol. 40: 4126-30). All samples were also subjected to concentration-floatation, sucrose gradient centrifugation and Baermann techniques for the detection of other endoparasites. A statistical analysis evaluating the prevalence of infection and possible risk factors (i.e. age, sex, breed, housing, presence of other parasites and chronic gastrointestinal signs) associated with T. foetus infection was performed using Fisher’s exact test and the odds ratio (OR). RESULTS AND CONCLUSIONS. Tritrichomonas foetus was detected in 18/105 (17.14%) cats. This infection rate is higher if compared to previous studies carried out in Italy (0.75 - 32.4%). In only two cats, T. foetus was found in association respectively with Giardia and A. abstrusus. The infection was detected more frequently in cats with chronic gastrointestinal signs (38.9% vs 14.9%; p = 0.01; 95% CI = 1.40-13.99; OR = 4.27) and no other statistically significant associations emerged. This study confirms the endemicity of T. foetus in cats from central Italy and its importance as a cause of chronic gastrointestinal signs. Moreover, the data here presented showed that the infection may also occur in adult and mixed-breed catsas previously reported (Doi et al., 2012J Vet Med Sci. 74: 413-41; Gruffydd-Jones et al., 2013 J Feline Med Surg. 15: 647-49)

Effect of vaccination against Glasser's Disease in a farm suffering from polyserositis in weaned pigs

Simple Summary This study mainly aims to investigate the impact and the etiology of swine polyserositis, focusing on three causative agents: Glaesserella parasuis, Streptococcus suis and Mycoplasma hyorhinis. Results indicate that the etiology of polyserositis is an intricate puzzle of pathogens, and that each pig herd likely represents a unique scenario. Therefore, the correct diagnosis of polyserositis can be really challenging and often makes the implementation of therapeutic and control strategies frustrating. Polyserositis mostly affects 4-8 weeks old piglets and is usually caused by Glaesserella parasuis, and/or Streptococcus suis, and/or Mycoplasma hyorhinis. The present study aimed to investigate the prevalence and etiology of polyserositis in a tricky pig herd. The concurrent effect of vaccination for Glasser's disease was also assessed. A total of 46 sows and 387 piglets were herein investigated, subdivided into three groups based on their immune status (i.e., vaccination of sows and piglets). All the piglets found spontaneously dead between the 2nd and 16th week of age were recorded and necropsied. Whenever polyserositis was diagnosed, biomolecular investigations were carried out to detect the above-mentioned pathogens. Mycoplasma hyorhinis was detected most frequently (n = 23), often as the only causative agent (n = 15), whereas S. suis was observed in 8 cases (6 as the only pathogen). Moreover, Glaesserella parasuis was demonstrated in 6 piglets, always in combination with Mycoplasma hyorhinis and/or Streptococcus suis. Vaccination did not significantly affect mortality rates. Overall, our data indicate that polyserositis is likely caused by an intricate puzzle of pathogens, even when dealing with a small herd and during a short time span. That makes it challenging to achieve the correct diagnosis and to properly manage this health issue

Electrochemical genosensors for the detection of Bonamia parasite. Selection of single strand-DNA (ssDNA) probes by simulation of the secondary structure folding

A post-PCR nucleic acid work by comparing experimental data, from electrochemical genosensors, andbioinformatics data, derived from the simulation of the secondary structure folding and prediction ofhybridisation reaction, was carried out in order to rationalize the selection of ssDNA probes for thedetection of two Bonamia species, B. exitiosa and B. ostreae, parasites of Ostrea edulis.Six ssDNA probes (from 11 to 25 bases in length, 2 thiolated and 4 biotinylated) were selected withindifferent regions of B. ostreae and B. exitiosa PCR amplicons (300 and 304 bases, respectively) with the aimto discriminate between these parasite species. ssDNA amplicons and probes were analyzed separatelyusing the “Mfold Web Server” simulating the secondary structure folding behaviour. The hybridisation ofamplicon–probe was predicted by means of “Dinamelt Web Server”. The results were evaluated consideringthe number of hydrogen bonds broken and formed in the simulated folding and hybridisation process,variance in gaps for each sequence and number of available bases. In the experimental part, thermallydenatured PCR products were captured at the sensor interface via sandwich hybridisation with surfacetetheredprobes (thiolated probes) and biotinylated signalling probes. A convergence between analyticalsignals and simulated results was observed, indicating the possibility to use bioinformatic data for ssDNAprobes selection to be incorporated in genosensors.[...

Short- and long-term effects of a dietary yeast ß-glucan (Macrogard) and alginic acid (Ergosan) preparation on immune response in sea bass (Dicentrarchus labrax).

The present study investigated the immunomodulatory activity of Ergosan, an algal extract containing alginic acid, and Macrogard, a yeast extract containing b-glucans, on innate and specific immunity in sea bass (Dicentrarchus labrax). Four cycles of experimental feeding using normal fish feed formulation (control group) supplemented with Ergosan (0.5%) or Macrogard (0.1%) were performed at 60-day intervals (15 days of treatment + 45 days of suspension). Serum complement, lysozyme, total proteins and heat shock protein (HSP) concentrations were measured at 15, 30 and 45 days from the end of the first 15-day feeding cycle (short term) and 45 days after the end of each feeding cycle over a 35-week period (long term). The percentage of B- and T-lymphocytes in peripheral blood leucocytes and gut were measured over long-term trial. Significant elevation (P<0.05) in serum complement activity occurred in sea bass fed with alginic acid and glucans, at 15 days from the end of first cycle of treatment. Significant elevation (P<0.05) in serum lysozyme, gill and liver HSP concentration were observed in the same experimental groups at 30 days from the end of treatment, whereas a significant increase (P<0.05) of complement activity was only observed in fish that received an Ergosan diet. At 45 days from the end of treatment, complement, lysozyme and HSP concentration did not differ among groups. Over the long-term period, no significant differences were observed in innate and specific immune parameters, survival, growth performances and conversion index in treated and control fish. A dramatic decrease of both innate and acquired immune parameters was observed during the winter season in all groups, followed by a partial recovery when water temperature increased. Reduction in complement and lysozyme activities was significatively correlated (P<0.01) to water temperature variation. The results suggested the potential of alginic acid and b-glucans to activate some innate immune responses in sea bass, and particularly under conditions of immunodepression related to environmental stress

Defensive response of European sea bass (Dicentrarchus labrax) against Listonella anguillarum or Photobacterium damselae subsp. piscicida experimental infection.

10Sea bass were experimentally infected with Listonella anguillarum or Photobacterium damselae subsp. piscicida (Phdp). At 24 and 72h post-infection, the expression analysis of immune-relevant genes (IL-1beta, IL-6, IL-8, Hepcidin), the transcriptional level and detection of HSP70, and the quantification of serum iron were investigated in association with the histological analysis and the bacterial recognition in tissues by immunohistochemistry. At 15 days post-infection, the specific antibody response was detected in surviving fish, as well as the transcriptional levels of TcR and BcR sequences. Both experimental infections were characterized by a similar acute response, whereas different histological and immunohistochemistry evidences were observed. In particular, the early reaction appeared suitable for the clearance of L. anguillarum, thus limiting the histological lesions, the bacterial dissemination and the further development of acquired immunity in surviving fish. On the contrary, the innate response appeared not enough to resolve the Phdp infection, which was characterized by tissue damage, bacterial widespread and substantial detection of specific humoral immunity in surviving fish, also associated to lymphocytes clonal expansion. Besides the opportunistic conditions involved in fish vibriosis and pasteurellosis, the comparison between these experimental infection models seems to suggest that the rate of development of the acquired immunity is strictly linked to the activation of the host innate response combined to the degree of bacterial virulence.reservedmixedMosca, F.; Ciulli, S.; Volpatti, D.; Romano, N.; Volpe, E.; Bulfon, C.; Massimini, M; Caccia, E.; Galeotti, M.; Tiscar, P.G.Mosca, F.; Ciulli, S.; Volpatti, Donatella; Romano, N.; Volpe, E.; Bulfon, Chiara; Massimini, M; Caccia, E.; Galeotti, Marco; Tiscar, P. G

Heat Shock Proteins (HSPs) 27, 72 and 73 in Normal and Pre-ulcerative Mucosa of the Gastric Pars Oesophagea in Swine

Heat shock proteins (HSPs), known to play a key role in cellular homeostasis, may also play a role in the defensive mechanisms of gastric mucosa. By means of appropriate immunohistochemical and immunobiochemical techniques, the expression of HSP27, HSP72 and HSP73 within the epithelium of normal and pre-ulcerative (hyperkeratinized) mucosa of the pars oesophagea of abattoir pigs was assessed. In normal mucosa, HSP72 and HSP73 expression was mainly limited to the basal epithelial cell layer, whereas HSP27 expression was consistently detected within the superficial epithelial cell layers. In hyperkeratinized mucosa, HSP72 and HSP73 immunoreactivity appeared to be more widespread, becoming very intense within epithelial cells affected by hydropic degeneration. Hyperkeratinized mucosa also showed HSP27 immunoreactivity, which was particularly intense in epithelial areas affected by hydropic degeneration. Western blot analysis confirmed HSP27, HSP72 and HSP73 expression in normal and in pre-ulcerative mucosa of the pars oesophagea. Semi-quantitative analysis showed that for all three HSPs the immunoreactivity was more intense in pre-ulcerative mucosa than in normal mucosa. The different expression patterns observed may have functional significance; further studies are needed, however, to define the role of HSPs in swine oesophagogastric lesions, the aetiology and pathogenesis of which are largely unknown.[...

Severe outbreak of pasteurellosis in sows: a case description

Sow mortality represents a relevant issue in the modern pig industry and can negatively impact the profitability of farming. Sow mortality rate should range between 3-6% on a yearly basis, but higher rates are quite commonly reported. Disease conditions of the locomotor, digestive, reproductive and urinary systems are most frequently observed in dead/euthanized sows. On the other hand, respiratory diseases are rare in this category of animals, the most severe episodes resulting from the introduction of a "new" infectious agent into the breeding stock, thus in absence of an adequate level of herd immunity. We describe herein the main clinical, pathological and microbiological findings observed during a severe outbreak of respiratory disease, which occurred in a breeding pig herd in Central Italy. The respiratory syndrome affected a very high percentage of pregnant and lactating sows (around 30%), both primiparous and multiparous. Within two weeks, the mortality rate reached 15%. At necropsy, serous-fibrinous pleuritis, along with large foci of acute bronchopneumonia, affecting the cranio-ventral parts of both lungs, represented the most relevant pathological findings in animals under study. In depth bacteriological investigations identified a capsular type D and hgbB-positive Pasteurella multocida strain as the sole causative agent. Pneumonic pasteurellosis usually affects grower/finishing pigs and represents the final stage of the so-called porcine respiratory disease complex. As a matter of fact, Pasteurella multocida rarely acts as the primary cause of porcine pneumonia, rather being considered an opportunist which complicates primary infections caused by other bacteria and/or viruses. The co-factors which likely contributed to determine such a severe respiratory syndrome in sows are largely unknown. Speculatively, we hypothesize that resulted from the introduction of a "new" strain of P. multocida, which infected immunologically naive sows. The poor level of biosecurity adopted, as well as the kinetics of this outbreak, argue in favour of such a scenario